Jessica Karen Wong/Notebook/2007-6-28

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

• 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