Jessica Karen Wong/Notebook/2007-7-11

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Colony PCR Gel from 7/11/07
Colony PCR Gel from 7/11/07
Gradient PCR Gel from 7/11/07
Gradient PCR Gel from 7/11/07


Contents

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
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