Jessica Karen Wong/Notebook/2007-7-11
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I2057
- 1AK3
- Overnight plate had tons of very small colonies - jason said it's probably not right
- Colony PCR'ed 4 samples
- Ran on gel - didn't show (the middle 4 lanes on top)
- Spun down rest of transformation and replated
- 3K3
- Overnight plate had many colonies that looked good
- PCRed 4 samples
- Ran on gel - looks the right size (1st 4 lanes on top after ladder)
- Made overnights to prep for sequencing
I2056
- Saw no growth on both I2056-1AT3 and I2056-1AC3 plates
- Spun down rest of transformation and plated
- Got sequencing back - still not built
I2055
- Got sequencing in for the religation
- Only matches at 110 - aka still doesn't contain the promoter
- Redid the colony PCRs (scarred product w/ pcr'ed in promoter in 1AK3)
- Used vent and a 1:30 ext time
- Comparing with a positive control of P1010 in 3K3
E0240
- Overnight plate of E0240-3K3 had a large number of colonies
- Colony PCRed 4 samples
- Ran on gel - all look to small
- Made overnights to prep for sequencing
- Got old E0240-3K3 sequencing back
- was messed up but only part there matched some of E0040 (GFP)
- Ran overnight BB PCR of E0240-1AK3 on a gel
- Of samples that showed up (we ran out of DNA) they were very smeary
- The only bright band on the gel is from our pos control P1010
- RePCR'ed overnight with Phusion at 53.5 w/ 1 min ext time
- Got sequencing of E0240-1AK3 back
- Sequencing was a bit messed up, but the Nsi/Pst site was scarred
- Sending for sequencing again just to make sure but Most Likely Scarred!
T9002
- Good haul on both overnight plates T9002-1AK3 and T9002-3K3
- Col PCR 4 samples of each
- Ran gel (bottom row) - all products were too small
- Made overnights to sequence
- Got sequencing back for T9002-3K3
- Sequence only matched some of the 3K3 plasmid
