User:Tara K. Luckau/Notebook/Team ConGen/Entry Base: Difference between revisions
From OpenWetWare
No edit summary |
|||
Line 94: | Line 94: | ||
* 270mL TAE + 4.05g agarose | * 270mL TAE + 4.05g agarose | ||
* (for future reference, total volume ≈ volume of gel box in cm<sup>3</sup>, ie 27cm long x 20 cm wide x 0.5 cm thick gel = 270mL of TAE) | * (for future reference, total volume ≈ volume of gel box in cm<sup>3</sup>, ie 27cm long x 20 cm wide x 0.5 cm thick gel = 270mL of TAE) | ||
==25 October 2010 - Ladder Mix== | |||
===Manufacturer specifications=== | |||
* 0.1µg ladder / mm width of well | |||
* STK at 1.0µg/µL | |||
===Other Knowns=== | |||
* well width (24-tooth comb) = 6mm | |||
* Jeremy (Bohonak lab) uses 5µL PCR product + 1.5µL dye | |||
===Calculations=== | |||
* 6mm x (0.1µg ladder)/(1mm width) x (1µL/1µg) = 0.6µL ladder per well | |||
* Per well: 0.6µL ladder + 4.4µL H<sub>2</sub>O + 1.5µL loading dye = 6.5µL total ladder mix | |||
* (loading dye = xylene cyanol and bromophenol, made by Bohonak lab) | |||
===Make Ladder Mix=== | |||
* 120µL ladder + 880µL H<sub>2</sub>O + 300µL loading dye = 1300µL ladder mix | |||
==25 October 2010 - Gel Scun2 F-R== | ==25 October 2010 - Gel Scun2 F-R== | ||
===Cast Gel=== | ===Cast Gel=== | ||
Line 125: | Line 140: | ||
* Next: | * Next: | ||
** try a bunch of different trials of ladder, gel stain, on small gel | ** try a bunch of different trials of ladder, gel stain, on small gel | ||
==26 October 2010 - GelRed info== | ==26 October 2010 - GelRed info== |
Revision as of 14:53, 26 October 2010
Tara's Lab Notebook | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page |
27 September 2010 - Ordered Primers
11 October 2010 - PCR Thermalcycler TestingPurpose
Results
Purpose
Results
12 October 2010 - PCR Thermalcycler TestingPurpose
Results
18 October 2010 - PCR Thermalcycler TestingPurpose
Results
18 October 2010 - Primer Rehydration, Scun2Purpose
Calculations
18 October 2010 - dNTP MixPurpose
Calculations
Wet Work
19 October 2010 - PCR Scun2 F-R21 October 2010 - Gel Scun2 F-RMake 1x TAE
Cast Gel
Next Try
25 October 2010 - Ladder MixManufacturer specifications
Other Knowns
Calculations
Make Ladder Mix
25 October 2010 - Gel Scun2 F-RCast Gel
Load GelRun Gel
Image GelWhat Happened?
26 October 2010 - GelRed info
|