Jessica Karen Wong/Notebook/2007-7-11

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