Jessica Karen Wong/Notebook/2007-7-30

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  • Sent for seq: all promoter testers cut w/ not1 (I2057 and E0240 S&X versions)
  • Diluted BB_R_Rev_Eco, BB_R_Rev_Spe, and BB_I2056_Fwd_X primers each to 40uM
  • BB PCRed both samples of I2056 to make both Eco and Spe versions (4 100ul rxns, 3:50ext time)
  • 10ul PCRed both preps of I2055 and the cell suspension of Col #1 with GFP internal Rev primer and each VF and VR
    • Checking to see if part is flipped, if flipped GFP_Rev and VR will show, if not GFP_Rev and VF will show
  • Testing Top10 competent cells by transforming and plating puc18
  • Made overnights of promoters J23116 and J23100 to miniprep, digest, and then test using E0240 and I2057
  • Transforming E0240-3K3-E/S, E0240-3K3-E/X, I2057-3K3-E/S, I2057-3K3-E/X with Knight Lab's DB3.1 electrocompetent cells
  • Ran a gel of today's PCRs loaded: I2056 Col #1 E, I2056-2-E, I2056-1-S, I2056-2-S, sp lad sp, I2055-3K3 GR &VF (1, 2, 1 cell), sp, I2055-3K3 GR&VR (1, 2, 1 cell)

Gel from 7-30