Jessica Karen Wong/Notebook/2007-6-28

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Contents

To Do

  • Redo PCR's of E0240, I2055, and colony
  • Re-digest B0032
  • Re-transform B0032 from Igem Distribution Kit
  • Replate yesterdays' transformations
  • Analyze the PCR gels
  • ligate and transform new devices?

PCR

Gel from 6/28/07
Gel from 6/28/07
  • Diluted dNTP's to 2.5uM
  • Did a 10 ul colony PCR on the 7 colonies of blue C
  • Did a 100ul preparatory PCR of E0240 at 49.5
  • PCR'ed I2055 on a gradient (12 10ul)
  • Ran a gel - Top is I2055, bottom Left is colony and bottom Right is E0240
    • E0240 has a very bright band at the right size
    • Colony PCR's didn't show
    • I2055 looks hazy but there might be something there?
  • PCR cleaned E0240

B0032

  • Digested the B0032 stock on 1A3 (in the measurement kit box) with EcoR1 and Spe1
    • DNA concentration was very low 45.2 ng/ul - hopfully that's ok
  • In case that batch is bad we transformed the Igem Distribution of B0032 to grow it up

RBS Tester

  • Yesterday's overnight transformation plates had no growth yet again
  • Spun down and replated the rest of yesterday's transformation
  • Ligated B0032-J04650-1AT3 and B-J-1AC3 from today's digest of B0032
  • Transformed today's ligation and plated for overnight
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