User:Steven J. Koch/Notebook/Kochlab/2009/03/29/Looking at old unzipping data/Example of all journal files
0001.ini
JUNK FILE TO SAVE SETTINGS
(segment000)
0002.ini
If using "Velocity Clamp New" then X wiggles will be subtracted (Wiggle file noted in journal) and Y will be replaced with the value of the actual wiggles subtracted (although averaged)
Protocol (1:30 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5 minutes; 5 s.v. PBS; 1 s.v. DNA about 5 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 10 minutes; 5 s.v. Dopes; seal
ROOM FAN OFF SINCE ABOUT 2 AM or earlier
(segment-specific comments below)
(segment000) Peeled partially then stopped (OLD)
(segment001) Peeled some, but stuck (NEW)
(segment002) Peeled a long way (NEW)...was there a geometry problem? I was fairly high
(segment003) Broke (NEW)
(segment004) Broke after a short time (OLD)
(segment005) Broke
(segment006) Peeled long way (OLD)...Now will look at data
0003.ini
If using "Velocity Clamp New" then X wiggles will be subtracted (Wiggle file noted in journal) and Y will be replaced with the value of the actual wiggles subtracted (although averaged)
Protocol (1:30 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5 minutes; 5 s.v. PBS; 1 s.v. DNA about 5 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 10 minutes; 5 s.v. Dopes; seal
ROOM FAN OFF SINCE ABOUT 2 AM or earlier
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) Awesome...think geometry was OK, peeled the entire way. Fluctuations were very evident and repeatable on oscilloscope, but not sure if i decimated this file too much (NEW Velocity Clamp)
(segment001) (OLD velocity clamp) Brok maybe 1/3 of the way through? COOL: When trap reset to center, it sucked bead back, and tether reannealed! So, will try to peel again in next segment
(segment002) (NEW Velocity clamp--doh! should have used old!) same as last segment after re-annealing
0004.ini
If using "Velocity Clamp New" then X wiggles will be subtracted (Wiggle file noted in journal) and Y will be replaced with the value of the actual wiggles subtracted (although averaged)
Protocol (1:30 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5 minutes; 5 s.v. PBS; 1 s.v. DNA about 5 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 10 minutes; 5 s.v. Dopes; seal
ROOM FAN OFF SINCE ABOUT 2 AM or earlier
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) A very huge file with only 10 decimation...I want Fluctuations!
0005.ini
Zeroed detector at 1.1 V (+/- 10 bits at gain 2 is the drift)
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
ROOM FAN OFF SINCE ABOUT 8 PM
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) Good peeler, I think (although it's tough to see distance when stretching with piezo. I check zero afterwards, and X was about -7.5 +/- 2.5 bits on gain 2. Switching to 1.5 volts drops the signal to -82.5 + / 2.5 bits...so i should have zeroed at higher voltage, damn
(segment001) This one peeled a lot longer than the previous one.
0006.ini
Zeroed detector at 1.1 V (+/- 10 bits at gain 2 is the drift)
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
ROOM FAN OFF SINCE ABOUT 8 PM
I think the "find tether center" did not work because I didn't load a wiggle array. Fuck.
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) JUNK FILE
0007.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) 10 decimation, set reference votlage to 1V
0008.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
(segment000) AFTER this segment, flowed in BsaI 1:50 in Dopes Buffer (flowed in about 3:29 AM) (I think a huge bubble must have washed across the surface, because the sample is almost entirely bare (compared to the normal amount of stuck beads previously)
(segment001) hmmm weird...seemed VERY long
(segment002) same as last segment...junk tether...skipping along the surface
(segment003) YES! Looked like a good restriction popper!
(segment004) darn...think i was too close to surface...it stuck
(segment005) broke right away
(segment006) Very weird. Strangely there was no popping of enzymes...until the very end. It is getting very warm in the room, so maybe that has something to do with it. I wonder if the enzyme was sliding along until it encountered the very end of the DNA? Actually, it is not THAT warm in the room...at least not as warm as the enzyme likes, anyway.
0009.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus, and then JUST slightly higher to ensure no sticking..then turn on trap (which brings bed up a little to be parfocal)
(segment-specific comments below)
SAME SAMPLE AS LAST FILE! I ACCIDENTALLY WROTE THE DATA
(segment000) this one also has no enzymes until the last part
(segment001) Now switched to the Naked DNA sample
0010.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus some of the time...not that exact...
(segment-specific comments below)
(segment000) Weird...still a tether when I was finished...will restretch in next segment
(segment001) Focused a bit higher...broke fairly soon.
(segment002) Looked nice and smooth...I need to change the program a little
0011.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus some of the time...not that exact...
(segment-specific comments below)
(segment000) Also fairly smooth, like last file
(segment001) Smooth (these don't seem to be overstretching...
(segment002) Weird...still a tether after overstretch
(segment003) Whoah! STILL a tether (even looks like same place!)
(segment004) OK...NOW the tether center is about 2 microns away
(segment005) weird...OK...I'm going to stop
0012.ini
Zeroed detector at 2 V
Protocol (1:30 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 10 s.v. Dopes;4 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. PBS; 1 s.v. Bangs 1:10 in PBS about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
(same sample...hours old)
ROOM FAN OFF SINCE ABOUT 1:45 AM
Focusing method: Focus up until bead is in focus some of the time...not that exact...
(segment-specific comments below)
(segment000)
(segment001) weird still a tether at end.
(segment002) looked good until the end...when it had a different tether center
(segment003) once again, had reformed a tether when I let go.
0013.ini
Zeroed detector at 1.4 V
Protocol (1:10 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
This tethering protocol worked better than the last time (at least for the 17-mer sample). I don't know if it's because of 3x higher DNA concentration or because of the DOPES buffer. 17-mer tether density is at least 1 per 90 micron, probably 3-4
Room fan off at 11:10 AM
Focusing method: on surface (using z-10)
(segment-specific comments below)
(segment000) broke early
0014.ini
Zeroed detector at 1.4 V
Protocol (1:10 dilution of 17-mer peeling template, or 1:10 dilution of stored PCR), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5-10 minutes; 2 s.v. BGB about 10 minutes; 5 s.v. PBS; 1 s.v. DNA about 10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only the PCR sample
This tethering protocol worked better than the last time (at least for the 17-mer sample). I don't know if it's because of 3x higher DNA concentration or because of the DOPES buffer. 17-mer tether density is at least 1 per 90 micron, probably 3-4
Room fan on, and noisy people in the room
Focusing method: z-8.5 method (focus on bead, just ever so slightly above tethered bead)
(segment-specific comments below)
(segment000) z-11 method
(segment001) z-11 method
(segment002)
(segment003) z-8.5 method
0015.ini
Zeroed detector at 1.4 V
Protocol (1:20 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5-10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only one chamber
Room fan on.
Focusing method: z-8.5 method (focus on bead, just ever so slightly above tethered bead)
(segment-specific comments below)
(segment000) first peeling try
0016.ini
Zeroed detector at 1.1 V
Protocol (1:20 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5-10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only one chamber
Room fan on.
Focusing method: z-8.5 method (focus on bead, just ever so slightly above tethered bead)
(segment-specific comments below)
(segment000) before flowing in
(segment001) about 30-50 seconds after flowing in 50 ul of ice cold dopes buffer. i bet it warmed up quite a bit, though
(segment002) many minutes later (probably fully warm again)
0017.ini
Zeroed detector at 1.1 V
Protocol (1:20 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5-10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only one chamber
Room fan on.
Focusing method: z-8.5 method (focus on bead, just ever so slightly above tethered bead)
(segment-specific comments below)
(segment000) trying force clamp within peeling
(segment001) same tether
0018.ini
Zeroed detector at 1.1 V
Protocol (1:20 dilution of 17-mer peeling template), 5 s.v. Dopes; 5 s.v. PBS; 2 s.v. antidig about 5 minutes; 2 s.v. BGB about 5-10 minutes; 5 s.v. PBS; 1 s.v. DNA about 5-10 minutes; 5 s.v. DOPES; 1 s.v. Bangs 1:10 in DOPES about 20-25 minutes; 10 s.v. Dopes; seal only one chamber
Room fan on.
Focusing method: z-8.5 method (focus on bead, just ever so slightly above tethered bead)
(segment-specific comments below)
(segment000)
(segment001) successfully sat on a opening / closing event. drift is evident, though
0019.ini
Junk File
(segment-specific comments below)
(segment000) dsDNA 12 bit board
0020.ini
dsDNA file (with offset subtraction)
(segment-specific comments below)
(segment000)
(segment001)
0021.ini
dsDNA file (with offset subtraction)
(segment-specific comments below)
(segment000) good one?
(segment001) good one?
(segment002)
(segment003)
0022.ini
dsDNA file (with offset subtraction)
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) overstretch?
0023.ini
dsDNA file (with offset subtraction)
(segment-specific comments below)
(segment000)
(segment001)
(segment002) very long "overstretch"
(segment003) broke at onset of overstretch?
0024.ini
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
0025.ini
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010) This started to peel during FTC
(segment011) Same tether as previous segment. So far, this is the only 0.35 um tether I could get that peeled.
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
0026.ini
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) Example of peeler in BsaIBuffer. This is the first tether I tried is said buffer.
0027.ini
This file is 0.53 micron beads, in BsaIBuffer, but NO enzyme.
I have failed to load a wiggle array.
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) I had previously peeled this tether, so will have some hysteresis. It is not good data, but shows artifactual stuff that COULD look like BsaI if I am not careful. I don't know what it is, buy my best guess is that it's multiply tethers.
(segment001)
(segment002) Another that shows artifacts
0028.ini
This file is 0.53 micron beads, in BsaIBuffer, but NO enzyme.
I have loaded most recent wiggle array.
z-telescope is at 10.5mm and I am trying to focus on surface?
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) example of peeler, geometry looked bad
(segment001) example of BAD things that can happen. Looks like peel / overstretch combo.
(segment002) broke
0029.ini
This file is 0.53 micron beads, in BsaIBuffer, but NO enzyme.
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) multiple tether
(segment005)
(segment006)
(segment007)
(segment008) shows artifactual (probably to close to surface)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013) peeled partially, and will now add BsaI
0030.ini
This file is 0.53 micron beads, in BsaIBuffer, ENZYME added at about 2:14
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) oops...forgot to turn laser on
(segment001) same tether as last segment of last file...shows at least 6 popping events
(segment002)
(segment003)
(segment004)
(segment005) dust in trap...only one pop
(segment006)
(segment007)
(segment008)
(segment009)
(segment010) welded
(segment011)
(segment012) ridiculous
(segment013) same as last
(segment014)
(segment015) welded
0031.ini
This file is 0.53 micron beads, in BsaIBuffer, ENZYME added at about 2:14
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : dopes ; 5.000000 s.v.; 1.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB 4 mg / ml ; 2.000000 s.v.; 20.000000 min.
Step 3 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer peeler, 1:30 dilution ; 1.000000 s.v.; 15.000000 min.
Step 5 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 6 : Beads* ; 1.000000 s.v.; 25.000000 min.
Step 7 : *Beads are either 0.35 dopes, or 0.53 BsaIBuf ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) no enzyme until the end
(segment001)
(segment002)
(segment003)
(segment004)
(segment005) VERY strange
(segment006)
(segment007)
(segment008) strange hiccup at the beginning, doesn't look right
(segment009) hmm...no enzyme
0032-small.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS NON_TWEEN
THIS SAMPLE SUCKS!
-------------------------------------------------------------
Step 0 : Peel Buffer + Tween ; 5.000000 s.v.; 0.000000 min.
Step 1 : Peel Buffer ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in PeelBuf ; 1.000000 s.v.; 11.000000 min.
Step 3 : BGB (old aliquots) ; 1.000000 s.v.; 5.000000 min.
Step 4 : BGB (repeat) ; 1.000000 s.v.; 5.000000 min.
Step 5 : PeelBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : 17-mer (most recent batch, 1:10 in PBS, 1:3 in PeelBuf) ; 1.000000 s.v.; 10.000000 min.
Step 7 : EcoRI Bufferf* ; 5.000000 s.v.; 0.000000 min.
Step 8 : Beads, Bangs 0.53 micron streptavidin, 1:10 into EcoRI Buffer* ; 1.000000 s.v.; 20.000000 min.
Step 9 : EcoRI Buffer* ; 10.000000 s.v.; 0.000000 min.
Step 10 : *1 sample with tween, the other without ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) junk
(segment001) junk
0032.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS NON_TWEEN
THIS SAMPLE SUCKS!
-------------------------------------------------------------
Step 0 : Peel Buffer + Tween ; 5.000000 s.v.; 0.000000 min.
Step 1 : Peel Buffer ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in PeelBuf ; 1.000000 s.v.; 11.000000 min.
Step 3 : BGB (old aliquots) ; 1.000000 s.v.; 5.000000 min.
Step 4 : BGB (repeat) ; 1.000000 s.v.; 5.000000 min.
Step 5 : PeelBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : 17-mer (most recent batch, 1:10 in PBS, 1:3 in PeelBuf) ; 1.000000 s.v.; 10.000000 min.
Step 7 : EcoRI Bufferf* ; 5.000000 s.v.; 0.000000 min.
Step 8 : Beads, Bangs 0.53 micron streptavidin, 1:10 into EcoRI Buffer* ; 1.000000 s.v.; 20.000000 min.
Step 9 : EcoRI Buffer* ; 10.000000 s.v.; 0.000000 min.
Step 10 : *1 sample with tween, the other without ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) junk
(segment001) junk
0033.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS TWEEN
-------------------------------------------------------------
Step 0 : Peel Buffer + Tween ; 5.000000 s.v.; 0.000000 min.
Step 1 : Peel Buffer ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in PeelBuf ; 1.000000 s.v.; 11.000000 min.
Step 3 : BGB (old aliquots) ; 1.000000 s.v.; 5.000000 min.
Step 4 : BGB (repeat) ; 1.000000 s.v.; 5.000000 min.
Step 5 : PeelBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : 17-mer (most recent batch, 1:10 in PBS, 1:3 in PeelBuf) ; 1.000000 s.v.; 10.000000 min.
Step 7 : EcoRI Bufferf* ; 5.000000 s.v.; 0.000000 min.
Step 8 : Beads, Bangs 0.53 micron streptavidin, 1:10 into EcoRI Buffer* ; 1.000000 s.v.; 20.000000 min.
Step 9 : EcoRI Buffer* ; 10.000000 s.v.; 0.000000 min.
Step 10 : *1 sample with tween, the other without ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) Peels at first but then weird!
(segment001) another fucked up one (was short amplitude at first)
(segment002)
0034.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000)peel z-8.85 actually for this segment. first attempt at peeling with this sample
(segment001) peel
(segment002) nope
(segment003) peel
(segment004) mistake
(segment005) i'm glad this one did this, because it looked much too long at first.
(segment006) broke soon (video)
(segment007) broke soon (video)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
0035-small.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000)
(segment001) 9 or 10 pops!
(segment002) 5 pops
(segment003) lots-o-pops
(segment004)
(segment005)
(segment006) 6 pops...2nd or 3rd one may have been funky
(segment007) VERY WEIRD...as if it got very long for some reason part way through
(segment008)
(segment009) 11 pops...5th one was like i said in segment 006...that is, instead of abrupt breakage, may have been almost slip-like
(segment010)
0035.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000)
(segment001) 9 or 10 pops!
(segment002) 5 pops
(segment003) lots-o-pops
(segment004)
(segment005)
(segment006) 6 pops...2nd or 3rd one may have been funky
(segment007) VERY WEIRD...as if it got very long for some reason part way through
(segment008)
(segment009) 11 pops...5th one was like i said in segment 006...that is, instead of abrupt breakage, may have been almost slip-like
(segment010)
0036.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) Still popping after all this time!!!
0036_more.ini
0037.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) normal speed popper
(segment001)
(segment002)
(segment003)
(segment004)
(segment005) good extremely fast one!
(segment006) good extremely fast one!
0038.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) STRANGE? did it slip?
(segment001) broke early
(segment002) very long overstretch?
(segment003)
(segment004) Holy crap that took a long time. popped a bunch of times. forces all over the place
0038Seg0004-DFSTweak.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) STRANGE? did it slip?
(segment001) broke early
(segment002) very long overstretch?
(segment003)
(segment004) Holy crap that took a long time. popped a bunch of times. forces all over the place
0039.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000)
(segment001) fast pop?
0040.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) same tether as last segment of last file
(segment001)
(segment002)
(segment003)
(segment004) awesome!
(segment005)
(segment006) another one with pops
(segment007) probably pops
(segment008)
(segment009) pops
(segment010)
(segment011) pops
0041.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS DOPES
(segment-specific comments below)
(segment000) broke immediately
(segment002) broke immediately
(segment003) broke immediately
(segment004) broke immediately
(segment005) broke after slight peeling
(segment006) broke immediately
(segment007) broke immediately
(segment008) peeled
(segment009) broke?
(segment010) broke immediately
(segment011) peeled maybe 1/2 way
(segment012) broke
(segment013) peeled
0042.ini
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) peeled full
(segment001) peeled a lot
(segment002) peeled a lot
(segment003) peeled a lot
0043.ini
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) no peel
(segment001) peel
(segment002) no peel
(segment003) forget
(segment004) peel
(segment005) peel
(segment006) no peel
(segment007) no peel
(segment008) peel
0044.ini
5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) peel
(segment001) peel
(segment002) peel
(segment003) peel
(segment004) peel
0045.ini
10 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) peel
(segment001) didn't see
(segment002) huh?
(segment003) peel
(segment004) peel part
(segment005) no
(segment006) no
(segment007) no
(segment008) no
(segment009) peel tiny bit
0046.ini
20 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) no
(segment001) no
(segment002) no
(segment003) peel
(segment004) peel partial
0047.ini
50 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) no
(segment001) no
(segment002) no
(segment003) peel
(segment004) no
(segment005) no
(segment006) no
(segment007) peel a bit
0048.ini
50 MHz / sec, FREE BEAD!!! TO ESTIMATE DRAG
(Not sure if configured correctly for this file
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) free bead
(segment001) free bead
(segment002) free bead
0049.ini
50 MHz / sec, FREE BEAD!!! TO ESTIMATE DRAG
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
(segment001)
0050.ini
50 MHz / sec, NO BEAD!!!
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
0051.ini
0.5 MHz / sec, NO BEAD!!!
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
0052.ini
50 MHz / sec, NO BEAD!!!
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
0053.ini
50 MHz / sec, FREE BEAD!!!
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
0054.ini
50 MHz / sec, NO BEAD!!!
FIXED OFFSET?
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
0055.ini
50 MHz / sec, FREE BEAD!!!
FIXED OFFSET?
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000)
(segment001)
0056.ini
EcoRI
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA + EcoRI
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) lots-o-popping, baby!
(segment001) popping
(segment002) 4 pops
(segment003) junk
(segment004) 3 or 4 pops
(segment005) no
(segment006) no
(segment007) 6 pops
(segment008) 7 pops
(segment009) 8 pops
(segment010) 4 pops
0057.ini
EcoRI
5MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA + EcoRI
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) no
(segment001) bad tether
(segment002) 3 pops
(segment003) no
(segment004) no
(segment005) at least 3 pops
(segment006) no
(segment007) maybe 1 pop
(segment008) no
(segment009) 4 pops
0058.ini
EcoRI
10MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA + EcoRI
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) some pops
(segment001) pop (stuff at end is a different stuck bead)
(segment002) no
(segment003) 1 pop
(segment004) no
(segment005) at least two pops
(segment006) 1 pop
0059.ini
EcoRI
20MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA + EcoRI
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) pop?
(segment001) at least 1 pop
(segment002) no
(segment003) peel no pop
(segment004) at least 2 pops
0060.ini
EcoRI
20MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
THIS SAMPLE IS 0.9 dopes + 10 mM EDTA + EcoRI
-------------------------------------------------------------
Step 0 : Dopes ; 1.000000 s.v.; 10.000000 min.
Step 1 : 0.9xPBS 10 mM EDTA ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig 20 ug / ml in 0.9xPBS + 10 mM EDTA ; 1.000000 s.v.; 11.000000 min.
Step 3 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 9.000000 min.
Step 4 : 10 mg / ml BGB in 0.45x PBS 5 mM EDTA ; 2.000000 s.v.; 5.000000 min.
Step 5 : 17-mer 1:11 in 10/11 PBS + 10 mM EDTA ; 1.000000 s.v.; 14.000000 min.
Step 6 : Bangs 0.53 streptavidin 1:10 in final buffer 0.81 PBS 9 mM EDTA ; 1.000000 s.v.; 15.000000 min.
Step 7 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
Step 8 : 0.9x Dopes + 10 mM EDTA ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
Have washed the second time now, but still no enzymes
(segment-specific comments below)
(segment000) no sure
(segment001) pops
(segment002) peel no pops
(segment003) pop
(segment004) no sure
0061.ini
Naked DNA
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) great peeler
0062.ini
Naked DNA
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) peel a little then overstretch.
(segment002) jhunk
(segment003) broke
(segment004) complete peel
(segment005) broek
(segment006) peel a little then freaky (styukc)(
(segment007) peeled about 1/2
(segment008) double tether (almost surely...double force, neighboring breakage, etc.)
(segment009) stuck
(segment010) peeled completely
(segment011) peeled completely
(segment012) peeled about 1/2
(segment013) broek
(segment014) broek
(segment015) peeled completely
0063.ini
EcorRI
0.5 MHz / sec
NOTE: Sample probably chilly at first. The overstretching inthe first segment is evidence of this.
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) awesome 11 popper (note: temperatuer probably cold at first!)
(segment001) awesome 13 popper
(segment002) 4 popper 1/2 peeler
(segment003) broek
(segment004) broke
(segment005) 4 popper 1/4 peeler
(segment006) 9 popper 3/4 peelr
(segment007) broke
(segment008) broek
(segment009) broke
(segment010) 6 popper full peeler?
(segment011) 10 popper
0064.ini
EcoRI
5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) couple pops
(segment001) broke
(segment002) 4 pops, couple overstretches
(segment003) broke
(segment004) 4 pops couple overstretches
(segment005) 6 pops, lots of overstretching
0065.ini
EcoRI
0.05 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) broke
(segment002) weird
(segment003) broke
(segment004) weird
(segment005) broke
(segment006) broke
(segment007) fucking broke
(segment008) 4 pops, narrow force distribution
(segment009) peel one pop
(segment010) lotsof pops, but unfortunately strange, probably won't be able to tell whether it was one tether or two somehow in parallel.
(segment011) 2 pops
(segment012) broke
(segment013) 11 pops, probably full peeler. there is a double popper in here (that is, seemes like two sites right next to each other).
(segment014) 12 pops...good to compare with segmetn 13
(segment015) sparse 7 pops
0066.ini
EcoRI
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled maybe full, only 1 pop
(segment001) broke
(segment002) 6 pops
(segment003) 10 pops
(segment004) broke
(segment005) broke
(segment006) 4 pops with an overstretch
(segment007) broke
0067.ini
EcoRI (freshly added)
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
Step 12 : EcoRI, 1:100 (same as last batch) at about 3:04 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) 12 pops
(segment001) 10 pops
(segment002) 14 pops
(segment003) 1 pop, short peel
(segment004) broke
(segment005) broke
(segment006) broke
(segment007) 10 pops
0068.ini
EcoRI (freshly added)
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
Step 12 : EcoRI, 1:100 (same as last batch) at about 3:04 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
000: broke
(segment001) at least 17 pops, overstrethc, too...oops, held foot down too long
0069.ini
EcoRI (freshly added)
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
Step 12 : EcoRI, 1:100 (same as last batch) at about 3:04 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) 1 pop
(segment002) broke
(segment003) 3 pops
(segment004) broke
(segment005) broke
(segment006) 4 pops
(segment007) 6 pops
(segment008) broke
0070.ini
EcoRI added, but now in empty buffer
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer(04/03/2001) ; 5.000000 s.v.; 0.000000 min.
Step 1 : Repeat ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig (Roche) 20 ug / ml diluted in Popping buffer ; 1.000000 s.v.; 10.000000 min.
Step 3 : 10 mg / ml BGB in 0.5 PBS + 5 mM EDTA ; 1.000000 s.v.; 10.000000 min.
Step 4 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 5 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 6 : 1:10 17-mer (+1mM EDTA today) in PBS ; 1.000000 s.v.; 10.000000 min.
Step 7 : Popping Buffer ; 5.000000 s.v.; 0.000000 min.
Step 8 : Bangs 0.53 1:10 into Popping Buffer ; 1.000000 s.v.; 20.000000 min.
Step 9 : Popping Buffer (about 1AM) ; 10.000000 s.v.; 0.000000 min.
Step 10 : delay ; 0.000000 s.v.; 83.000000 min.
Step 11 : EcoRI, 1:100 into Popping Buffer (2:23 AM) ; 5.000000 s.v.; 0.000000 min.
Step 12 : EcoRI, 1:100 (same as last batch) at about 3:04 AM ; 5.000000 s.v.; 0.000000 min.
Step 13 : 1:5 EcoRI into popping buffer 3:14 AM ; 2.000000 s.v.; 0.000000 min.
Step 14 : Plain Popping Buffer (no enzyme) 3:22AM, ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) 1 pop
(segment001) 12 pops
(segment002)
(segment003) 1 pop
(segment004) 3 pops
(segment005) 11 pops
(segment006) weird (probably transient sticking)
0071.ini
BsaI
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer, room temp ; 5.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml (1:10 in popping buffer) ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB, same as yesterday ; 2.000000 s.v.; 10.000000 min.
Step 3 : Pop Buffer ; 5.000000 s.v.; 0.000000 min.
Step 4 : BGB ; 2.000000 s.v.; 10.000000 min.
Step 5 : 17-mer +EDTA (see yesterday) ; 1.000000 s.v.; 10.000000 min.
Step 6 : Pop Buffer ; 10.000000 s.v.; 0.000000 min.
Step 7 : Bangs 0.53 streptavidin 1:10 into Pop Buffer (Diluted yesterday...when flowing in, the color is non-homogenous, probably indicating clumping from the buffer?) ; 1.000000 s.v.; 10.000000 min.
Step 8 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 9 : Pop Buff ; 10.000000 s.v.; 0.000000 min.
Step 10 : BsaI 1:25 into Popping Buffer ; 1.000000 s.v.; 2.000000 min.
Step 11 : Repeat above 5 times (ended about midnight) ; 0.000000 s.v.; 0.000000 min.
Step 12 : seal nail polish ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pops
(segment001) broke
(segment002) broke
(segment003) pops
(segment004) broek FTC
(segment005) MANY pops
(segment006) broke
(segment007) broke
(segment008) few pps
(segment009) probably all the pops
(segment010)
(segment011) probably complete pops
(segment012) pops
0072.ini
BsaI
0.5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer, room temp ; 5.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml (1:10 in popping buffer) ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB, same as yesterday ; 2.000000 s.v.; 10.000000 min.
Step 3 : Pop Buffer ; 5.000000 s.v.; 0.000000 min.
Step 4 : BGB ; 2.000000 s.v.; 10.000000 min.
Step 5 : 17-mer +EDTA (see yesterday) ; 1.000000 s.v.; 10.000000 min.
Step 6 : Pop Buffer ; 10.000000 s.v.; 0.000000 min.
Step 7 : Bangs 0.53 streptavidin 1:10 into Pop Buffer (Diluted yesterday...when flowing in, the color is non-homogenous, probably indicating clumping from the buffer?) ; 1.000000 s.v.; 10.000000 min.
Step 8 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 9 : Pop Buff ; 10.000000 s.v.; 0.000000 min.
Step 10 : BsaI 1:25 into Popping Buffer ; 1.000000 s.v.; 2.000000 min.
Step 11 : Repeat above 5 times (ended about midnight) ; 0.000000 s.v.; 0.000000 min.
Step 12 : seal nail polish ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pops
(segment001) partial pops
(segment002) broke
(segment003) pops NOTE: THIS ONE SHOWS THE PROBLEMATIC RISE IN FORCE
(segment004) partial pops
(segment005) crazy
(segment006) pops
(segment007) broke
(segment008) broke
(segment009) pops
0073.ini
BsaI
5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer, room temp ; 5.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml (1:10 in popping buffer) ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB, same as yesterday ; 2.000000 s.v.; 10.000000 min.
Step 3 : Pop Buffer ; 5.000000 s.v.; 0.000000 min.
Step 4 : BGB ; 2.000000 s.v.; 10.000000 min.
Step 5 : 17-mer +EDTA (see yesterday) ; 1.000000 s.v.; 10.000000 min.
Step 6 : Pop Buffer ; 10.000000 s.v.; 0.000000 min.
Step 7 : Bangs 0.53 streptavidin 1:10 into Pop Buffer (Diluted yesterday...when flowing in, the color is non-homogenous, probably indicating clumping from the buffer?) ; 1.000000 s.v.; 10.000000 min.
Step 8 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 9 : Pop Buff ; 10.000000 s.v.; 0.000000 min.
Step 10 : BsaI 1:25 into Popping Buffer ; 1.000000 s.v.; 2.000000 min.
Step 11 : Repeat above 5 times (ended about midnight) ; 0.000000 s.v.; 0.000000 min.
Step 12 : seal nail polish ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) WOW probably all the pops
(segment001) broke
(segment002) partial pop
(segment003) partial pop
(segment004) lots o pops
(segment005) lots o pops
(segment006) lots o pops, but for some reason overstretch (dual tether?)
(segment007) pops, but again overstretch
(segment008) partial pops
0074.ini
BsaI
5 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer, room temp ; 5.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml (1:10 in popping buffer) ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB, same as yesterday ; 2.000000 s.v.; 10.000000 min.
Step 3 : Pop Buffer ; 5.000000 s.v.; 0.000000 min.
Step 4 : BGB ; 2.000000 s.v.; 10.000000 min.
Step 5 : 17-mer +EDTA (see yesterday) ; 1.000000 s.v.; 10.000000 min.
Step 6 : Pop Buffer ; 10.000000 s.v.; 0.000000 min.
Step 7 : Bangs 0.53 streptavidin 1:10 into Pop Buffer (Diluted yesterday...when flowing in, the color is non-homogenous, probably indicating clumping from the buffer?) ; 1.000000 s.v.; 10.000000 min.
Step 8 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 9 : Pop Buff ; 10.000000 s.v.; 0.000000 min.
Step 10 : BsaI 1:25 into Popping Buffer ; 1.000000 s.v.; 2.000000 min.
Step 11 : Repeat above 5 times (ended about midnight) ; 0.000000 s.v.; 0.000000 min.
Step 12 : seal nail polish ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) partial pop
(segment001) no pops?
(segment002) pops, but overstretch
(segment003) partial pops
(segment004) very strange
(segment005) lots o pops
0075.ini
BsaI
10 MHz / sec
I have loaded most recent wiggle array.
z-telescope is at now at 8.5, and I will use the focusing method where I make bead parfocal, and then just ever so slightly raise above that
-------------------------------------------------------------
Step 0 : Popping Buffer, room temp ; 5.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml (1:10 in popping buffer) ; 2.000000 s.v.; 10.000000 min.
Step 2 : BGB, same as yesterday ; 2.000000 s.v.; 10.000000 min.
Step 3 : Pop Buffer ; 5.000000 s.v.; 0.000000 min.
Step 4 : BGB ; 2.000000 s.v.; 10.000000 min.
Step 5 : 17-mer +EDTA (see yesterday) ; 1.000000 s.v.; 10.000000 min.
Step 6 : Pop Buffer ; 10.000000 s.v.; 0.000000 min.
Step 7 : Bangs 0.53 streptavidin 1:10 into Pop Buffer (Diluted yesterday...when flowing in, the color is non-homogenous, probably indicating clumping from the buffer?) ; 1.000000 s.v.; 10.000000 min.
Step 8 : repeat ; 1.000000 s.v.; 10.000000 min.
Step 9 : Pop Buff ; 10.000000 s.v.; 0.000000 min.
Step 10 : BsaI 1:25 into Popping Buffer ; 1.000000 s.v.; 2.000000 min.
Step 11 : Repeat above 5 times (ended about midnight) ; 0.000000 s.v.; 0.000000 min.
Step 12 : seal nail polish ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) not sure
(segment001) broke
(segment002) broek
(segment003) pops
(segment004) crazy
(segment005) complete popper
(segment006) complete popper
0076.ini
Richard's Template...See 2001/04/16 notes in his directories
This file with My most recent wiggle array (like april 05)
(segment-specific comments below)
(segment000) breaks at overstretch (the most common event)
0077.ini
Richard's Template...See 2001/04/16 notes in his directories
This file with My most recent wiggle array (like april 05)
(segment-specific comments below)
(segment000)
0078.ini
Richard's Template...See 2001/04/16 notes in his directories
This file with My most recent wiggle array (like april 05)
(segment-specific comments below)
(segment000)
0079.ini
See following data files for protocol, sorry
(segment-specific comments below)
(segment000) broke
(segment001)
(segment002) peeled a little
(segment003) peeled a little
(segment004) peeled a little
(segment005)
(segment006) started as stuck
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
0080.ini
EcoRI
0.5 Mhz / s
trying to focus on surface, but using eyepiece
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) Yes! I think a pop!
(segment001) stuck
(segment002) BAD DATA SET...a bead was in the trap during offset array
(segment003) broke
(segment004) broke
(segment005) broke
(segment006) peel, but no pop
(segment007) broke
(segment008) stuck
0081.ini
EcoRI
0.5 Mhz / s
pRL574 template
giving up focusing on surface...using bead method again
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled, popped, but maybe in wrong place?
(segment001) peeled, broke early
(segment002) junk
(segment003) broke
(segment004) broke
(segment005) peeled, no pop
(segment006) peeled, no pop
(segment007) junk
(segment008) bad
(segment009) broke
0082.ini
EcoRI
0.5 Mhz / s
pRL574 template
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled, no pop! (added Eco after this segment)
(segment001) peeled no pop!
(segment002) broke
(segment003) peeled, broke before pop
(segment004) funky
(segment005)non-specific end-to-end tether
(segment006) double peeler
(segment007) peeled, popped. HOWEVER, bead was near trap during offset
(segment008) double peeler
(segment009) peeled, no pop
(segment010) bad, i think
(segment011) peeled, no pop
0083.ini
EcoRI
5 Mhz / s
pRL574 template
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled, no pop
(segment001) junk
(segment002) broke
(segment003) non-specific teth
0084.ini
EcoRI, 1:5
0.55 Mhz / s
pRL574 template
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 26.000000 min.
Step 11 : EcoRI, 1:5 in PopBuf (LifeTech) (about 12:30PM) ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled and popped...was a little bit of junk in the trap
(segment001) peeled and popped
(segment002) peeled, broke early
(segment003) peeled, broke early
(segment004) peeled and popped!
(segment005) peeled and popped!
0085.ini
EcoRI, 1:5
5 Mhz / s
pRL574 template
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 26.000000 min.
Step 11 : EcoRI, 1:5 in PopBuf (LifeTech) (about 12:30PM) ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) definitely a pop!...maybe more (also has freaky reversible shit!)
(segment001) Holy shit! some kind of really freaky reversible fluctuation going on!!!
(segment002) junk
(segment003) messed up
(segment004) messed up
(segment005) pop
(segment006) broke
(segment007) pop
(segment008)
(segment009) peel no pop
(segment010) bad
(segment011) pop (not at canonical)
(segment012) peel no pop?
(segment013) pop, but not at canonical (same site as 11)
0086.ini
0.5 Mhz / s
4.4 kb double tagged
back to surface focusing method, using video to fucus
zeroed at 1.5
IGNORE THE ECORI STEPS BELOW!!!
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 26.000000 min.
Step 11 : EcoRI, 1:5 in PopBuf (LifeTech) (about 12:30PM) ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) broke
(segment002) bead was interacting during offset
(segment003) broke
(segment004) broke
(segment005) broke
(segment006) broke
0087.ini
0.5 Mhz / s
Added EcoRI about 30 seconds before first segment
4.4 kb double tagged
back to surface focusing method, using video to fucus
zeroed at 1.5
IGNORE THE ECORI STEPS BELOW!!!
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 26.000000 min.
Step 11 : EcoRI, 1:5 in PopBuf (LifeTech) (about 12:30PM) ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) mysteriously overstretches now?
(segment001) broke
(segment002) broke
(segment003) broke
(segment004) broke
(segment005) overstretch
(segment006) overstrech
0088.ini
0.5 Mhz / s
For this file, I flowed in 2 sample volumes of 2 Molal Sucrose in 50% PoppingBuffer
Added EcoRI during last file.
4.4 kb double tagged
back to surface focusing method, using video to fucus
zeroed at 1.5
IGNORE THE ECORI STEPS BELOW!!!
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : antidig (Roche) 20 ug / ml in PBS ; 2.000000 s.v.; 7.000000 min.
Step 3 : BGB 10 mg / ml in ? (same as last time) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : DNA [either pRL574 peeling template from december, or 17 pM dsDNA PCR) ; 1.000000 s.v.; 14.000000 min.
Step 6 : Dopes (ran out of thawed popBuf) ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf 0.45 micron streptavidin, freshly diluted ; 1.000000 s.v.; 22.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 40.000000 min.
Step 9 : EcoRI, 1:50 in PopBuf (LifeTech) (about 11:43PM) ; 2.000000 s.v.; 20.000000 min.
Step 10 : EcoRI, 1:50 in PopBuf (LifeTech) (about 12:04PM) ; 2.000000 s.v.; 26.000000 min.
Step 11 : EcoRI, 1:5 in PopBuf (LifeTech) (about 12:30PM) ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) overstretch!
(segment001) overstretch
(segment002) overstretch
(segment003) broke
(segment004) broke
(segment005) partial overstretch
(segment006) double force overstretch
(segment007) suppressed overstretch or two in parallel
(segment008) overstretch
(segment009) bit of an overstretch
(segment010) broke
(segment011) bad
(segment012) broke
(segment013) complete overstretch
0089.ini
0.5 Mhz / s
Naked 17-mer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) good peel
(segment001) broke
(segment002) good peel
(segment003) bad
(segment004) broke
(segment005) broke
(segment006) broke
(segment007) complete peel
0090.ini
0.5 Mhz / s
Naked 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) broke
(segment002) peeled!!!
(segment003) non-specific overstretch
(segment004) mistake
(segment005) peeled!
0091.ini
0.05 Mhz / s
Naked 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled...fluctuations are evident...I wonder if their time scale is different, though?
(segment001) broke
(segment002) peeled strange...the end got pinned
(segment003) broke
(segment004) complete peel
0092.ini
0.05 Mhz / s
Naked 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled
(segment001) messed up (may be large file)
(segment002) messed up
(segment003) broke
(segment004) broke
(segment005) peeled
0093.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) was this is??
(segment002) also strange
(segment003) bad
(segment004) bad
(segment005) broke
(segment006) kind of like 1...am I seeing it???
(segment007) overstretch
(segment008) broke
(segment009) missed
(segment010) overstretch
(segment011) overstretch
(segment012) overstretch
0094.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) whoa momma! the mother load!!!
(segment001) like 000
(segment002) still overstretch
0095.ini
0.005 Mhz / s
BsaI 1:20 17-mer IN 2 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke?
(segment001) peeled and pops all over the place?
(segment002) whoa...a long time
0096.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 1 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) popping, but less overstretching than with 2 m Sucrose
(segment001) pop
(segment002) overstretch?
(segment003) overstrretch again
(segment004) bad
(segment005) broke
(segment006) this one more like 001 and 000
0097.ini
0.005 Mhz / s
BsaI 1:20 17-mer IN 1 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) some kind of popping...didn't seem much different than the 2 m Sucrose stuff
0098.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 1 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) overstretch
(segment001) overstretch
(segment002) overstretch
0099.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 0.5 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) overstretch
(segment002) popping! (on the verge of overstretch)
(segment003) broke
(segment004) popping!
0100.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 0.5 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke strangely
(segment001) not sure...some kind of popping
(segment002) maybe a couple pops
(segment003) broke
(segment004) some kind of popping and overstretch?
(segment005) broke
(segment006) still freaky high force crap
(segment007) GOOD ONE
0101.ini
0.005 Mhz / s
BsaI 1:20 17-mer IN 0.5 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peel, no pop
(segment001) probably i am seeing multiple DNA?>
(segment002) peeled a bit, maybe 1 pop
(segment003) multiple DNA again?
(segment004) pops
0102.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 0.250 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) popping and overstretch
(segment002) bad
(segment003) broke
(segment004) overstretch
(segment005) broke
(segment006) overstretch
(segment007) popping...I think this one is REAL
(segment008) bad
(segment009) overstretch
0103.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 0.250 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) complete peeler
0104.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 0.125 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) not sure
(segment001) maybe a pop
(segment002) overstretch
(segment003) weird
(segment004) hmmmm...is this real? popping, but STILL at extremely high force
0105.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 0.125 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeling hten maybe a pop
(segment001) hmmmm looked real at first
(segment002) i am still quite suspicious of peeling in parallel
0106.ini
0.5 Mhz / s
BsaI 1:20 17-mer IN 0.0625 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) overstretch
(segment001) overstretch
(segment002) crazy popping
(segment003) crazy popping
(segment004) non-sprecific
(segment005) overstretch
0107.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 0.0625 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) looks like fucking multiple tethers
(segment001) broke
(segment002) peeled a bit
(segment003)
(segment004) peeled a bit
(segment005)
(segment006) still looks crappy
(segment007)
010706_ssDNA_HindIIIFit.ini
010706_ssDNAParams.ini
0108.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN 0.0625 m SUCROSE BUFFER!
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) nothing
(segment001) weird
(segment002)
(segment003) pops
(segment004) pop?
(segment005) pop?
(segment006)broke
(segment007) broke
(segment008) pops
010830_ssDNAParams_HindIIICappedFit_StiffRatio2p1_height775nm.ini
0109.ini
0.05 Mhz / s
BsaI 1:20 17-mer IN popping buffer only (+ 1:20 BsaI)
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : antidig ; 2.000000 s.v.; 8.000000 min.
Step 2 : BGB (same tube that's been in fridge for weeks) 10 mg / ml ; 2.000000 s.v.; 10.000000 min.
Step 3 : 17-mer from -80 C (contains EDTA now) ; 1.000000 s.v.; 10.000000 min.
Step 4 : PopBuffer ; 10.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 23.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : 2 m Sucrose in 50% popBuf (at about 3:10 AM) ; 5.000000 s.v.; 0.000000 min.
Step 8 : 2 m Sucrose 45% popBuf, 1:20 BsaI at about 3:38 AM ; 5.000000 s.v.; 0.000000 min.
Step 9 : 1 m Sucrose 70% popBuf, 1:20 BsaI about 4:10 AM ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) crappy
(segment002) broke
(segment003)
(segment004) peeled a bit
(segment005) peeled a bit, perhaps popping
(segment006) peeled with popping, but too high forces (multiple template?)
(segment007)
(segment008) yeah, that's fucking exactly what it is: multiple templates
0110.ini
0.5 Mhz / s
Naked 17-mer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : wash slide with water and kim wipes ; 0.000000 s.v.; 0.000000 min.
Step 1 : popBuf ; 5.000000 s.v.; 0.000000 min.
Step 2 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 3 : antidig (from last night) 20 ug / ml PBS ; 1.000000 s.v.; 4.000000 min.
Step 4 : 10 mg / ml BGB (same tube on 4C for weeks) ; 2.000000 s.v.; 6.000000 min.
Step 5 : 17-mer, 1:5 dilution of fridge stock, which is probably about 34 pM, I believe ; 1.000000 s.v.; 6.000000 min.
Step 6 : PopBuf or [90% popBuf 400 mm Sucrose] ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in [90 % PopBuf] or [80% popBuf 400 mM Sucrose] ; 0.000000 s.v.; 25.000000 min.
Step 8 : PopBuf or [90% PopBuf 400 mM Sucrose] ; 10.000000 s.v.; 0.000000 min.
Step 9 : counted tethers and stretched in 1 data file only ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled, then strange
(segment001) broke
(segment002) b
(segment003) peeled then strange, just like 000
(segment004) good peeler
(segment005) b
(segment006) b
(segment007) b
(segment008) b
(segment009) half peeler
(segment010) SUCROSE this segment and below
(segment011) good peeler
(segment012) partial peeler
0111.ini
0.5 Mhz / s
Naked 17-mer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled partial
(segment001) junk
(segment002) b
(segment003) b
(segment004) peel / overstretch
(segment005) peel partial
(segment006) broke?
(segment007) complete peel
(segment008) stuck
(segment009) complete peel
(segment010) b
(segment011) weird
(segment012) b
(segment013) b
(segment014) complete peel
0112.ini
0.5 Mhz / s
bsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) popping AND overstretching...why???
(segment001) popping AND overstretching
(segment002) popping
(segment003) broke?
(segment004) b
(segment005) pop, broke early
(segment006) pops
0113.ini
0.05 Mhz / s
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) looked like beautiful popping
(segment001) partial peel
(segment002) shows mysteriously high popping forces
(segment003) very good, like 000, same forces, too
0114.ini
1.85 V Force Clamp
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) Very HUGE segment...I believe showed one FC pop...tether did not break, and will continue in next file
0115.ini
Force Clamp higher force
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) still did not break
0116.ini
2.1 V Force Clamp
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) ok! broke this time (NOTE, last three files segment000 all the same tether
(segment001) pops!
(segment002) bad
(segment003) awesome!
(segment004) b
(segment005) b
(segment006) 11 pops, I think
(segment007) a few fast pops
(segment008) 7 pops, then stuck for a while
(segment009) bad
(segment010) bad
(segment011) bad
(segment012) lots of pops
0117.ini
2.2 V FC
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) bad
(segment001) WOW lots of popping
(segment002) lots of pops, then stuck for a while
(segment003) bad
(segment004) strangely high force...maybe parallel
(segment005) brple
(segment006) b
(segment007) bad
(segment008) unbreakable
(segment009) maybe a pop
0118.ini
0.05 Mhz / s
BsaI 1:20 popping buffer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) This segment shows that sample is still loaded with BsaI (well, not really, but it means it's plausible)
0119.ini
0.5 Mhz / s (except first segment)
BsaI washed away with 5 s.v. PopBuf
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) wow...lot's of popping (this segment ONLY 0.05MHz / s)
(segment001) tons of popping
(segment002) funky
(segment003) 14 pops
(segment004) b
(segment005) ~14 pops
(segment006) broke early
(segment007) b
0120.ini
0.5 Mhz / s
BsaI washed away with another 5 s.v. PopBuf
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) b
(segment001) 4 pops (broke early or not?)
(segment002) 6 pops (broke early or not?)
(segment003) 8 pops
(segment004) 10 pops
0121.ini
0.5 Mhz / s
BsaI washed away with 15 s.v. PopBuf so far
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) 4 pops
(segment001) 5 pops, seem closely spaced
(segment002) tons of pops
(segment003) 10 pops
(segment004) b
(segment005) 3 pops broke early
(segment006) broke
(segment007) b
(segment008) b
(segment009) freaky
(segment010) probably two in parallel
0122.ini
0.5 Mhz / s
BsaI washed away with 20 s.v. PopBuf so far
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) STILL tons of popping
(segment001) broke early
(segment002) tons of popping
(segment003) complete popping...I give up for now
0123.ini
0.5 Mhz / s
BsaI washed away with 20 s.v. PopBuf so far
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) overstretch
(segment001) 5 pops then break (looks closely spaced)
(segment002) 3 pops then break
(segment003) looked rather saturated
(segment004) also still looked saturated! persistent little fuckers
0124.ini
0.5 Mhz / s
BsaI w1:20,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) strange
(segment001) overstretch
(segment002) broke
(segment003) bad
(segment004) broke
(segment005) bad
(segment006) hmmmmm!!! two right in a row only!!!
(segment007) yes! zero...i am in the right ball park!
(segment008) zero again
(segment009) zero again
(segment010) partial peel (zero)
(segment011) broke
0125.ini
0.05 Mhz / s
BsaI w1:20,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) not good news...two only and i am going very slowly
(segment001) broke
(segment002) peel, broke no pop
(segment003) b
(segment004) b
(segment005)
(segment006)
(segment007) no pops, just weird thing at the end
(segment008) accidentally took foot off pedal
(segment009) same as last (pre-peeled somewhat) peeled, no pops
(segment010) broke very early
0126.ini
0.05 Mhz / s (LAST SEGMENT IS AFTER ADDING 1:5,000)
BsaI w1:20,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peel, no pop, broke early
(segment001)
(segment002) peeled completely, no pop. I would say right now, that the probability of template being occupied is about 1/5. I will therefor add
(segment003)
0127.ini
0.05 Mhz / s
BsaI 1:5,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) strange...peels, then overstretch
(segment001) hmmmmmm...not sure if a pop or what
(segment002) peeled no pop
0128.ini
0.5 Mhz / s
BsaI 1:5,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) strange...overstretch at end?
(segment001) peel, no pop
(segment002)
(segment003) overstretch then break?
(segment004)
(segment005)
(segment006)
(segment007)
(segment008) was too long to begin with
(segment009) only at very end again
(segment010) peel, no pop
0129.ini
5 Mhz / s
BsaI 1:5,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002) peel, no pop
(segment003) strange
(segment004) peel, no pop
(segment005) peel, no pop
0130.ini
5 Mhz / s
BsaI 1:2,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pop?
(segment001)
(segment002) pop?
0131.ini
0.5 Mhz / s
BsaI 1:2,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peel, no pop, broke early
(segment001) peel, no pop
(segment002) junk
(segment003)
(segment004) pop, then break
(segment005)
(segment006) SINGLE POP
(segment007) peel no pop
(segment008) POP...possibly smaller one following
0132.ini
0.05 Mhz / s
BsaI 1:2,000 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) for some reason overstretch after some peeling
(segment004) bad
(segment005) peel no pop
(segment006) peel broke early
(segment007) peel no pop
0133.ini
0.05 Mhz / s
BsaI 1:2,000 in popBuff (2nd wash)
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) strange
(segment001) forgot to zero before this file
(segment002)
(segment003)
(segment004) peel no pop
0134.ini
0.5 Mhz / s
BsaI 1:500 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) single pop-break
(segment001)
(segment002)
(segment003) peel/overstretch
(segment004)
(segment005)
(segment006) hmmmm 3 pops
(segment007)
(segment008)
(segment009)
(segment010) oh yeah! 3 right in a row at the end!
(segment011) junky
(segment012)
(segment013) 5 pops, then overstretch
(segment014) a few pops, ends with overstretch
(segment015) 6 pops
(segment016) pop / overstretch
(segment017)
(segment018)
(segment019) 3 pops break early
(segment020)
(segment021)
(segment022) definitely more pops at the end of the template. But unfortunately, I think I have too many enzymes right now
0135.ini
0.05 Mhz / s
BsaI 1:500 in popBuff
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) fucking peel/overstretch
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011) peeled broke early
(segment012) peeled broke very early
(segment013)
(segment014) peeled and lots of pops
0136.ini
0.5 Mhz / s
Naked DNA
Suface focusing method
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
0137.ini
2.1 fc
Naked 17-mer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002) hmmm...force clamp sticks, but i don't have enzyme yet
(segment003)
(segment004) giving up on this sample
0138.ini
2.1 fc
Naked 17-mer
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) peeled completely
0139.ini
2.1 fc
BsaI 1:2,000
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) whoah, momma! lots of pops
(segment005) maybe a pop, maybe not
(segment006)
(segment007)
(segment008) would not pop
(segment009) same as last, still will not pop
(segment010) bad
(segment011) same as 008 and 009, will not pop
(segment012)
(segment013)
(segment014) pops
(segment015)
(segment016)
(segment017)
(segment018) at least two pops
(segment019) broke or naked?
(segment020) would not pop
(segment021) at least 3 pops
(segment022)
(segment023)
(segment024)
(segment025)
(segment026) would not pop
(segment027)
(segment028) many, many pops
(segment029)
(segment030)
(segment031) probably a double tether
(segment032) junk
(segment033)
(segment034)
(segment035)
(segment036)
(segment037)
(segment038) pops
0140.ini
2.1 fc
BsaI 1:2000
back to surface focusing method, using video to fucus
zeroed at 1.5
-------------------------------------------------------------
Step 0 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 1 : PBS ; 5.000000 s.v.; 0.000000 min.
Step 2 : Antidig from yesterday (20 ug / ml) ; 2.000000 s.v.; 5.000000 min.
Step 3 : BGB 10 mg / ml (same tube at +4C for weeks) ; 2.000000 s.v.; 10.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : 17-mer, 1:5 dilution of freezer stock in PopBuf ; 1.000000 s.v.; 12.000000 min.
Step 6 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 7 : Bangs 1:10 in PopBuf ; 1.000000 s.v.; 35.000000 min.
Step 8 : PopBuf ; 10.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) 6 pops
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006) popped, but didn't finish (held)
(segment007) single pop, broke
(segment008) junk
(segment009) single pop, broke
(segment010) popped, then held
0141.ini
Junk?>
(segment-specific comments below)
(segment000) same tether as last file, popping is gone...this is a good example of something, but don't know what
(segment001)
(segment002)
0142.ini
2.1 fc
naked 17-mer
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001)
0143.ini
0.5 MHz / s
Naked 17-mer
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001) peeled partial
(segment002) peeled complete
(segment003) b
(segment004) peeled complete
0144.ini
0.5 MHz / s
EcoRI 1:20,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) partial peel
(segment001) junk
(segment002) peel no pop
(segment003)
(segment004) peel no pop
(segment005) some kind of pop?
0145.ini
FC 2.1
EcoRI 1:20,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peeled
(segment001)
(segment002)
(segment003) peeled
(segment004) peeled, then FC complete, no pops
0146.ini
0.5 MHz /s
EcoRI 1:20,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peeled partial
(segment001)
(segment002)
(segment003) pop?
(segment004) pop?
(segment005) partial peel, no pop
(segment006)
(segment007) suspicious
(segment008) b
(segment009) too long
(segment010)
(segment011) peel no pop
(segment012) peel no pop
(segment013) pop?
(segment014) pop?
(segment015) whoah!!! was this it>? unfortunately, looked strange
0147.ini
0.5 MHz /s
EcoRI 1:20,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peel no pop
(segment001)
(segment002) pop, then freaky
(segment003)
(segment004) partial peel
(segment005) held ("held" means overstretch / break)
(segment006)
(segment007)
(segment008) peel no pop (geometry bad)
(segment009)
(segment010)
(segment011) peel no pop
(segment012)
(segment013) held
(segment014) peel no pop
(segment015)
(segment016) peel no pop
0148.ini
0.5 MHz /s
EcoRI 1:20,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001) peel no pop
(segment002) held
(segment003) peel no pop
(segment004)
(segment005) peel no pop
(segment006)
(segment007)
(segment008)
(segment009) held
(segment010) partial peel no po
(segment011) bad
(segment012) peel no pop
(segment013)
(segment014) strange
(segment015)
(segment016)
(segment017)
(segment018) peel no pop
0149.ini
0.5 MHz /s
EcoRI 1:5,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) held at end
(segment001) TWO DISTINCT POPS
(segment002) peel, no pop
(segment003)
(segment004)
(segment005)
(segment006) peel, no pop, strange at the beginnbing
(segment007) peel, no pop
(segment008) partial peel, no pop
(segment009)
(segment010)
(segment011) peel, no pop
0150.ini
FC 2.1
EcoRI 1:5,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peeled, no pop
(segment001) bad
(segment002) peeled completely, no pop
0151.ini
0.5 Mhz / s
EcoRI 1:5,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peel, no pop?
(segment001)
(segment002)
(segment003) peel no pop
(segment004) partial peel, no pop
(segment005)
(segment006) partial peel, no pop
(segment007) held / pop at end?
(segment008) partial peel, no po
(segment009)
(segment010)
(segment011) two pops / break
(segment012) peel no pop
(segment013)
(segment014) peel no ppop
(segment015)
0152.ini
0.5 Mhz / s
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000) peel, no pop
(segment001)
(segment002)
(segment003) single pop
(segment004) single pop
(segment005) peel no pop
(segment006) partial peel, no pop
(segment007) partial peel, no pop
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013) peel no pop
0153.ini
0.5 Mhz / s
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005) Held here
0154.ini
FC
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000) fresh tether...peel, no pop
(segment001) held
0155.ini
FC
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000) held
0156.ini
FC 2.5
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000) same tether as last file
(segment001) finally popped
0157.ini
FC 2.4
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
THERE IS DEBRIS IN THE IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001) peeled, no pop
0158.ini
0.5 MHz / s
EcoRI 1:2,000
bead focusing method
zeroed at 1.5
removed debris
(segment-specific comments below)
(segment000)
(segment001) very closely spaced pops
(segment002) partial peel, no pops
(segment003)
(segment004) peel, no pop
(segment005) too long to begin with
(segment006) held
(segment007) partial peel no po
(segment008) peel no pop
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014) partial peel no pop
0159.ini
0.5 MHz / s
EcoRI 1:1,000
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001) single pop at very beginning
(segment002) held
(segment003) like every fucking site???!@
(segment004) classic double tether (empty by the way)
(segment005) peel no pop!
(segment006) held
(segment007) partial peel nop p
(segment008) held
(segment009)
(segment010)
(segment011)
(segment012) peel no pop
(segment013) single pop / held
(segment014) partial, no pop
(segment015) peel no pop
(segment016)
(segment017) partial, no pop
(segment018)
(segment019)
(segment020) parital no pop
(segment021)
(segment022)
(segment023)
(segment024) peel no pop, kind of junky
(segment025)
(segment026) pop/break
(segment027) peel no pop
(segment028) OOOPS, this is the 1:500
(segment029)
(segment030) two pops at 1:500
0160.ini
0.5 MHz / s
EcoRI 1:500
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peel no pop
(segment001) pop/break
(segment002) peel no ppo
(segment003) held
(segment004)
(segment005) peel no pop
(segment006)
(segment007)
(segment008)
(segment009) partial peel no pop
(segment010)
(segment011) held
(segment012)
(segment013) peel no pop...maybe EcoRI is being sequestered?
0161.ini
0.05 MHz / s
EcoRI 1:500
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000) peel, no pop
(segment001) partial
(segment002)
(segment003)
(segment004) two pops at end?
(segment005) over
(segment006) probably a double at the beginning?
0162.ini
0.5 MHz / s
EcoRI 1:100
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001)
(segment002) 3 pops
(segment003)
(segment004) 5 pops
(segment005)
(segment006)
(segment007)
(segment008) 1 pop, break early
(segment009) bad one, cuz another stuck bead affected
(segment010)
(segment011) closely spaced pops?
(segment012)
(segment013)
(segment014) single pop
(segment015) break early
(segment016) 4 pops, break early
(segment017) 2 pops, break
(segment018)
(segment019) jukn
0163.ini
FC2.2. / Hold
EcoRI 1:100
bead focusing method
zeroed at 1.5
(segment-specific comments below)
(segment000)
(segment001)
(segment002) peeled then FC / hold
(segment003) same tether as last
(segment004) same as last
(segment005) same as last
0164.ini
naked DNA, zeroed at 1.5 V
(segment-specific comments below)
(segment000)
(segment001) funky
(segment002) peeled completely
0165.ini
naked 17-mer (new template)
0.05 MHz / sec
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pinned for no particular reason
(segment001) repeat of template, pinned again
(segment002) looks like a double tether
0166.ini
naked 17-mer (new template)
0.5 MHz / sec
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) double tether
0167.ini
naked 17-mer (new template)
0.5 MHz / sec
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pinned and broke
(segment001) bad
(segment002) off center
(segment003) good
(segment004) good (same focus as last segment)
(segment005) stuck
(segment006) complete peel
(segment007) stuck
(segment008) stuck
(segment009) strange at the beginning...I would say this is my best focusing yet
(segment010) incomplete peeler
(segment011)
(segment012) stuck
(segment013) peel
0168.ini
naked 17-mer (new template)
various speeds reversibility
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) good reverser for study
(segment001) repeat of last one
0169.ini
BsaI 17-mer (new template)
various speeds reversibility
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
5:47 AM, added 5 s.v. BsaI 1:50 in PopBuf
-------------------------------------------------------------
(segment-specific comments below)
(segment000) popped, then broke
(segment001) popped, then broke
(segment002) popped, then reversed
(segment003) repeated
(segment004) repeated, but broke and reformed tethering about 2 microns SW (southwest)
0170.ini
BsaI 17-mer (new template)
various speeds reversibility
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
5:47 AM, added 5 s.v. BsaI 1:50 in PopBuf
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) good one
(segment002) drifted away during create offset array
(segment003) broke before end
(segment004) reversed...looked strange, though?
(segment005) repeat and broke
(segment006)
(segment007) broke
(segment008)
(segment009)
(segment010)
(segment011) ok...probably another good one to look at
0171.ini
BsaI 17-mer...trying reversal before pop
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
5:47 AM, added 5 s.v. BsaI 1:50 in PopBuf
-------------------------------------------------------------
(segment-specific comments below)
(segment000) This is the same (already peeled) template as last file...but i think it worked
(segment001) bad
(segment002) broke
(segment003) worked
(segment004) repeated
(segment005) repeated
(segment006) repeated faster release
0172.ini
BsaI 17-mer...trying reversal before pop
Focusing on piece of dust, z - 11 mm
zeroed X&Y detector at 1.5 V
room fan is off
-------------------------------------------------------------
Step 0 : Begin with Bert's specially cleaned coverglass and slide, no further pre-rinsing ; 0.000000 s.v.; 0.000000 min.
Step 1 : Antidig 20 ug / ml in Popping Buffer ; 1.000000 s.v.; 5.000000 min.
Step 2 : BGB, 10 mg / ml in 50% popping buffer. Note, this BGB is maybe 1 or 2 months old at +4C...However, it has been left out at room temperature 2 or 3 times for over 24 hours each time. Oh well, I'm not going to remake it now. ; 5.000000 s.v.; 5.000000 min.
Step 3 : PopBuff ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer peeling template, estimated 29 pM (Diluted 1:10 from stock into Popping Buffer) ; 1.000000 s.v.; 8.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, freshly diluted 1:10 into popping buffer, and then sonicated (100 ul volume) for about 1 minutes by hand ; 1.000000 s.v.; 8.000000 min.
5:47 AM, added 5 s.v. BsaI 1:50 in PopBuf
-------------------------------------------------------------
(segment-specific comments below)
(segment000) good one
(segment001) broke
(segment002) broke
(segment003) broke
(segment004) strange
(segment005)
(segment006) broek
(segment007) probably a double tether
(segment008) borke
(segment009) broke
(segment010) probably good enugh...need to stop
0173.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-650nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-700nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-700nm-Temp1.01-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-750nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-775nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-800nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0173Seg0006-900nm-DFSTweak.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) complete, but no cap
(segment004)
(segment005)
(segment006) YES?
(segment007)
(segment008) complete, no cap
(segment009)
(segment010)
(segment011) complete no cap
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0174.ini
17-mer XhoI
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) early
(segment001) YES?
(segment002) repeat
(segment003) early
(segment004) early
(segment005) strange, but perhaps
(segment006) YES?
(segment007) early
(segment008) YES?
(segment009) repeat
(segment010) uncut?
(segment011) complete uncut
(segment012) YES
(segment013) YES
(segment014)
0175.ini
17-mer XhoI
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) same tether as last segment of last file
(segment001)
(segment002)
(segment003) double tether
(segment004)
(segment005)
0176.ini
17-mer XhoI
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) other bead interfered with offset array
(segment001) same as last, other bead more otu of way, but still may interfere a bit
(segment002) another good one
(segment003) bad
(segment004) broke before reversal
(segment005)
(segment006)
(segment007)
(segment008)
0177.ini
17-mer HindIII
Focusing on piece of dust, z - 11 mm (camera Auto Enhance is off)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) broke
(segment001) no cap
(segment002)
(segment003) no cap
(segment004) broke
(segment005)
(segment006) no cap
(segment007) stuckl
(segment008) double tether
(segment009)
(segment010)
(segment011) freaky double tether
(segment012)
(segment013)
(segment014)
(segment015)
(segment016) capped but broke before reversal
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023) capped in the wrong place
0177Seg 16-DFSTweak.ini
0178.ini
Anchoring segment only (so, should not peel) See following files for protocol
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
0179.ini
Now have added T4 DNA ligase control (no DNA)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) This one evidence of higher force
(segment005) also
(segment006) also this one
(segment007) stuck
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
0180.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) bad
(segment005)
(segment006)
(segment007) bead in trap during create offset array
(segment008) same here
(segment009)
(segment010) bad offset array
(segment011)
0181.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002) high force
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
0182.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) high force
(segment005) bad offset
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
0183.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002) bad off arr
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
0184.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
0185.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
0186.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) peel!
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006) long peeler
(segment007)
(segment008)
(segment009)
0187.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) overstretch, but did not break, will repeat in next segment, after having moved stage and refocused
(segment004) overstretch and broke
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
0188.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
0189.ini
Now have added entire reaction (7 PM)
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
(segment-specific comments below)
(segment000) complete peeler
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016) complete peeler, possibly DNA ligase at sticky end
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
0190.ini
Sample B (Will be P18-Cap-High LIgation attempt) this is pre-ligation reaction control
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008) HIGH FORCE
(segment009)
(segment010)
0191.ini
P18-Cap-High Ligation Attempt
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) stuck
(segment004)
(segment005) this one?
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012) bad array
(segment013)
(segment014)
(segment015)
0192.ini
Sample B, P18-cap-high ligation reaction
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001) Looked like peeling?
(segment002)
(segment003)
(segment004)
(segment005)
(segment006) this one?
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
0193.ini
Sample B, P18-cap-high ligation reaction
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) perhaps a peeler
(segment005)
0194.ini
Sample B, P18-cap-high ligation reaction
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001) hmmm
(segment002)
(segment003)
0195.ini
Back to sample A
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
0196.ini
Back to sample A
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
HAVE BUBBLES / DUST IN IMMERSION OIL
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) complete peeler
0197.ini
New sample, super-fast protocol.
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000) combination peeling and sticking
(segment001)
(segment002) partial peel
(segment003) partial peel
(segment004) peel combination stick (i must be too low!)
(segment005)
(segment006) stuck
(segment007) combination peel / stick
(segment008) stuck
0198.ini
New sample, super-fast protocol.
Focusing on piece of tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002) i would say peelingn without stickign
(segment003)
(segment004) yup...i thought was a bit too low to begin with, and it stuck
(segment005) incomplete peeler
(segment006)
(segment007)
0199.ini
New sample, super-fast protocol.
Focusing on tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
0200.ini
New sample, super-fast protocol.
Focusing on tethers (the old parfocal method, which I like better)
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000) partial peel no stick
(segment001) partial peel no stick
(segment002) pee/ stick, although at end, focus was clearly too low
(segment003)
(segment004) double tether / complete peel no stick
(segment005)
(segment006) peel / partial stick
(segment007)
(segment008) wow, z-11, stick when i thought i was on the surface focus
(segment009) z--11
(segment010) z-11
0201.ini
New sample, super-fast protocol.
FOCUSING METHOD: z - 10.25; focus by looking at laser reflection off coverglass; looking at top right part of image, and focusing till it "slows" down near a vertical kind of pattern. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail). Nomarski is adjusted so that 0.48 bangs beads have a small white dot on bottom left of bead...this is probably near the nomarksi setting which makes dust maximally visible).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007) funky tether
0202.ini
New sample, super-fast protocol.
FOCUSING METHOD: z - 10.25; focus by looking at laser reflection off coverglass; looking at top right part of image, and focusing till it "slows" down near a vertical kind of pattern. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail). Nomarski is adjusted so that 0.48 bangs beads have a small white dot on bottom left of bead...this is probably near the nomarksi setting which makes dust maximally visible).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001) OK
0203.ini
XhoI added
0.5 MHz / sec
(Notice the strong star site very early)
FOCUSING METHOD: z - 10.25; focus by looking at laser reflection off coverglass; looking at top right part of image, and focusing till it "slows" down near a vertical kind of pattern. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail). Nomarski is adjusted so that 0.48 bangs beads have a small white dot on bottom left of bead...this is probably near the nomarksi setting which makes dust maximally visible).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002) difficult to say whether a stick or a pop
(segment003) broke early
(segment004) broke early
(segment005) two pops
(segment006) two or two/3 pops
(segment007) pop / break
(segment008) not sure, maybe stuck
(segment009) broke early
(segment010) broke early
(segment011) pop, broke
(segment012) broke early
(segment013)
(segment014) pop/broke at early star site
(segment015) pop/broke at early star site
0204.ini
XhoI added
0.5 MHz / sec
(Notice the strong star site very early)
FOCUSING METHOD: z - 10.25; focus by looking at laser reflection off coverglass; looking at top right part of image, and focusing till it "slows" down near a vertical kind of pattern. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail). Nomarski is adjusted so that 0.48 bangs beads have a small white dot on bottom left of bead...this is probably near the nomarksi setting which makes dust maximally visible).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001) peel, maybe pop, broke early
(segment002) broke early
(segment003) broke early
(segment004) lots of binding
(segment005) a pop
(segment006) probably shows 3 out of the 4 sties
(segment007)
(segment008) single pop
0205.ini
Naked 17-mer
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000) freaky double tether
(segment001)
(segment002)
(segment003) bad offset array
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
0206.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0013-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0013-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0015-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0015-DFSTweak.ini
0206Seg0015-DFSTweak.ini.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0016-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0016-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0020-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0021-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0021-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0027-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0027-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0029-010913-XhoI-dsCappedStyle.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0206Seg0029-DFSTweak.ini
XhoI 17-mer, 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) what one would expect (3 pops)
(segment016) again
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028) DISCOUNT...tether stuck
(segment029)
(segment030)
0207.ini
XhoI 17-mer, 5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
0208.ini
XhoI 17-mer, 0.005 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002) full length
(segment003) broke immediate
(segment004) broke mid
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010) too big initially
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) double tether
(segment016)
(segment017)
0209.ini
XhoI 17-mer, 0.05 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000) double tether
(segment001)
(segment002) double
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024) DISCARD ...broke
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
(segment030)
0209Seg0005-DFSTweak.ini
0209Seg0005-DFSTweak.ini.ini
XhoI 17-mer, 0.05 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000) double tether
(segment001)
(segment002) double
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024) DISCARD ...broke
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
(segment030)
0210.ini
XhoI 17-mer, 0.005 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001) still has an enzyme
0211.ini
XhoI, washed many times with popping buffer 17-mer, 0.5 MHz / s
Still breaking a lot, even though i do not see popping
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
0212.ini
XhoI, washed many times with popping buffer 17-mer, 0.5 MHz / s
Still breaking a lot, even though i do not see popping
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
0213.ini
XhoI, washed many times with popping buffer 17-mer, 0.5 MHz / s
washed many times with 1 mM tris, and with 2 M NaCl Tween-20
Still breaking a lot, even though i do not see popping
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
SEEING EXTRAORDINARY NOISE ON BERGEN OSCILLOSCOPE
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
0214.ini
BamHI 0.5 Mhz / S
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
0215.ini
BamHI 0.5 Mhz / S
PLEASE NOTE THAT THERE IS EXTRANEOUS BINDING! Somehow seems related to the 6 & 12 mer problem!
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001) ******Has an extra even at the end!!!
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
0216.ini
BamHI 0.5 Mhz / s
With 10 mM CaCl
(EXCEPT LAST FEW SEGMENTS ARE OTHER SAMPLE, ECORV)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009) strange hump
(segment010)
(segment011)
(segment012) OOOPS FORGOT TO SWITCH FILES!
(segment013) strange (EcoRV)
(segment014) strange RV
(segment015) OK RV
0217.ini
EcoRV 0.5 Mhz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) looks normal
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
0218.ini
EcoRV 5 Mhz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015) double?
0219.ini
BamHI 0.05 Mhz / s
With 10 mM CaCl
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001) bad offset array
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
0220.ini
EcoRV 0.05 MHz / s
With 10 mM CaCl in PBS
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) popping
0221.ini
EcoRV 0.05 MHz / s
With 10 mM CaCl in PBS
There is Calcium Precipitated everywhere
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Start with Karen's (stolen from Richard, cleaned by Brent) slides...not so clean ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig 20 ug / ml in Pop Buf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB ; 2.000000 s.v.; 7.000000 min.
Step 3 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer, 1:2 (already 1:10 earlier) in Pop Buf, estimated 15 pM ; 1.000000 s.v.; 3.000000 min.
Step 5 : Pop Buf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10 Pop Buf, sonicated earlier today ; 1.000000 s.v.; 10.000000 min.
Step 7 : Pop Buf ; 10.000000 s.v.; 0.000000 min.
Step 8 : One sample only (for now) BamHI, 1:200 in Pop Buf (Life Tech) ; 0.000000 s.v.; 0.000000 min.
Step 9 : Stretch For a While ; 0.000000 s.v.; 0.000000 min.
Step 10 : Other Sample, EcoRV at about 4:34 AM (1:50 in PopBUf) ; 5.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
0222.ini
BsoBI 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003) Lots o pops
(segment004)
(segment005)
0223.ini
BsoBI 0.0005 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) pins somewhere in the middle...but not a bad segment to look at
0223Seg 0-DFSTweak.ini
0224.ini
BsoBI 0.5 MHz / s
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008) possibly shows sliding
0225.ini
BsoBI 5 MHz / s
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014) A little non-canonical blip?
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020) bad offset array
(segment021) non-canon blip?
(segment022)
(segment023)
(segment024) sliding?
(segment025)
(segment026)
(segment027)
(segment028)
(segment029) oops..let up footswitch
(segment030) same as last segment
0225Seg 2-DFSTweak.ini
0226.ini
BsoBI 5 MHz / s
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007)
(segment008)
(segment009)
(segment010)
(segment011)
(segment012)
(segment013)
(segment014)
(segment015)
(segment016)
(segment017)
(segment018)
(segment019)
(segment020)
(segment021)
(segment022)
(segment023)
(segment024)
(segment025)
(segment026)
(segment027)
(segment028)
(segment029)
0226Seg 22-DFSTweak.ini
0227.ini
BsoBI 5 MHz / s
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001)
(segment002)
(segment003)
(segment004) broke during reversal??
(segment005) broke during reversal??
(segment006)
0228.ini
BsoBI 5 MHz / s
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001) same
(segment002) same
(segment003) same
0229.ini
BsoBI 0.5 MHz / s
reveresal
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: z - 10.25; focusing on laser spot, calibrated previously by small dust...different than last time, because Z&Y telescope have changed. Camera details are probably important (Gain - 10; offset - 4.1; AGC on; no boost; maximum detail).
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Bert's slides ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh, 1:10 in PopBUf (20 ug / ml) ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB 10 mg / ml, ; 2.000000 s.v.; 10.000000 min.
Step 3 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 4 : 17-mer, 1:4 in PopBuf ; 1.000000 s.v.; 6.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs, soncicated, 1:10 PopBUf ; 1.000000 s.v.; 12.000000 min.
Step 7 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 8 : BsoBI (one sample only), 1:50 in PopBuf ; 2.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001) same (shows no pops)
(segment002)
(segment003)
(segment004)
(segment005)
(segment006)
(segment007) no pops in the first place
(segment008) no pops
(segment009)
(segment010)
(segment011)
0230.ini
Naked 17-mer in sealed chamber
z - 11 mm
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: Focusing on surface (smallest dust available)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 8.000000 min.
Step 3 : New 17-mer (1:4 dilution of previous 1:10 dilution...was also at room temperature last night) (Forgot to wash before this step) ; 1.000000 s.v.; 3.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : Seal ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) full peel
(segment001) full peel
(segment002) full peel
(segment003) full peel
(segment004) half peel
(segment005) full peel
0231.ini
Naked 17-mer in sealed chamber
z - 13 mm
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: Focused on a small piece of dust at z - 13.0 mm, and I have memorized (i hope) what the diffraction pattern reflection at voltage 1.0 should look like. [Video on Koch VII, 10:09 AM
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 8.000000 min.
Step 3 : New 17-mer (1:4 dilution of previous 1:10 dilution...was also at room temperature last night) (Forgot to wash before this step) ; 1.000000 s.v.; 3.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : Seal ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) full peel
(segment001) full peel
(segment002) broke
(segment003) at least close to full
(segment004) strange
(segment005) broke
(segment006) double then full, but should not use this one
(segment007) half peel
(segment008) broke
(segment009) bad
(segment010) bad
(segment011) broek
(segment012) broke
(segment013) weird, but full (don't use)
(segment014) full
0232.ini
Naked 17-mer in sealed chamber
z - 12 mm
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: Focused on a small piece of dust at z - 13.0 mm, and I have memorized (i hope) what the diffraction pattern reflection at voltage 1.0 should look like. [Video on Koch VII, 10:09 AM
Even though this file is z-12 mm, i still focus using laser when z - 13 mm. (Yes, this is a pain in the ass)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 8.000000 min.
Step 3 : New 17-mer (1:4 dilution of previous 1:10 dilution...was also at room temperature last night) (Forgot to wash before this step) ; 1.000000 s.v.; 3.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : Seal ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) broke
(segment001) close enough to full that i am counting it!
(segment002) full
(segment003) broke
(segment004) broke
(segment005) 1/2 peel
(segment006) full peel
(segment007) broke
(segment008) broke
(segment009) full peel, stick at the end
0233.ini
Naked 17-mer in sealed chamber
z - 11 mm
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: Focused on a small piece of dust at z - 13.0 mm, and I have memorized (i hope) what the diffraction pattern reflection at voltage 1.0 should look like. [Video on Koch VII, 10:09 AM
Even though this file is z-12 mm, i still focus using laser when z - 13 mm. (Yes, this is a pain in the ass)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 8.000000 min.
Step 3 : New 17-mer (1:4 dilution of previous 1:10 dilution...was also at room temperature last night) (Forgot to wash before this step) ; 1.000000 s.v.; 3.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : Seal ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) 1/4 peel
(segment001) broke
(segment002) broke
(segment003) double tether complete
(segment004) full peel
(segment005) broke
(segment006) broke
(segment007) triple tether and bad offset array
(segment008) badly off in y direction???
(segment009) tiny peel
(segment010) strange in middle (compare this to segment 24 of some file yesterday)
(segment011) bad
(segment012) long enough
0234.ini
Naked 17-mer in sealed chamber
z - 10.25 mm
This is sample B (1:10,000 BsoBI at 3:57 PM)
FOCUSING METHOD: Focused on a small piece of dust at z - 13.0 mm, and I have memorized (i hope) what the diffraction pattern reflection at voltage 1.0 should look like. [Video on Koch VII, 10:09 AM
Even though this file is z-xx mm, i'm still focus using laser when z - 13 mm. (Yes, this is a pain in the ass)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 8.000000 min.
Step 3 : New 17-mer (1:4 dilution of previous 1:10 dilution...was also at room temperature last night) (Forgot to wash before this step) ; 1.000000 s.v.; 3.000000 min.
Step 4 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 5 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 6 : PopBuf ; 10.000000 s.v.; 0.000000 min.
Step 7 : Seal ; 0.000000 s.v.; 0.000000 min.
-------------------------------------------------------------
(segment-specific comments below)
(segment000) tiny peel
(segment001) broke
(segment002) broke
(segment003) 1/2 peel, but enough
(segment004) stuck...interesting
(segment005) broke
(segment006) close enough for gov. work
0235.ini
Naked 17-mer
z - 10.25 mm
FOCUSING METHOD: Focused on a small piece of dust at z - 13.0 mm, and I have memorized (i hope) what the diffraction pattern reflection at voltage 1.0 should look like. [Video on Koch VII, 10:09 AM
Even though this file is z-xx mm, i'm still focus using laser when z - 13 mm. (Yes, this is a pain in the ass)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 1.000000 min.
Step 3 : BGB ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer ( no dilution of previous 1:10 dilution...was also at room temperature last night) ; 1.000000 s.v.; 3.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 7 : PopBuf, supplemented with 1 mM Biotin (which also results in 1% DMSO) ; 10.000000 s.v.; 0.000000 min.
Step 8 : wash with popbuf, 5 s..v
-------------------------------------------------------------
(segment-specific comments below)
(segment000)
(segment001) full peeler
(segment002) not stick, but strange
(segment003)
(segment004)
(segment005) partial
(segment006)
(segment007) strange...not stick? or maybe stick? but unstick?
(segment008) full peel
0236.ini
Naked 17-mer
z - 10.25 mm
FOCUSING METHOD: Focusing on beam at z - 10.25, accounting for shape difference compared with z - 13 mm (earlier today method)
zeroed X&Y detector at 1.5 V
room fan is ON
-------------------------------------------------------------
Step 0 : Karen's Slides & cover ; 0.000000 s.v.; 0.000000 min.
Step 1 : antidig, fresh 1:10 popBuf ; 1.000000 s.v.; 3.000000 min.
Step 2 : BGB (I left it at room temperature AGAIN!...probably like 6 hours) ; 2.000000 s.v.; 1.000000 min.
Step 3 : BGB ; 5.000000 s.v.; 0.000000 min.
Step 4 : New 17-mer ( no dilution of previous 1:10 dilution...was also at room temperature last night) ; 1.000000 s.v.; 3.000000 min.
Step 5 : PopBuf ; 5.000000 s.v.; 0.000000 min.
Step 6 : Bangs 1:10, freshly in popBuf, sonicated 1 minute ; 1.000000 s.v.; 10.000000 min.
Step 7 : PopBuf, supplemented with 1 mM Biotin (which also results in 1% DMSO) ; 10.000000 s.v.; 0.000000 min.
Step 8 : wash with popbuf, 5 s..v
-------------------------------------------------------------
(segment-specific comments below)
(segment000) Good one, but hiccups right in the middle!
(segment001)