Team:Brown/Notebook/August17

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Tuesday, August 17, 2010

Transformations:

  • bglII-CI-pGEM
  • bglII-AraC-pGEM
  • bglII-Mnt-pGEM
  • RFC control (in BL21s as we are running out of XL1Bs

Plated w/ 2% xGal, 2ul IPTG for blue white selection on Amp+ LB plates.

Gel

  1. Lab Ladder
  2. Wessel lab ladder
  3. LovTwo PCR

Gel 81710.jpg

Gel extracted lovtwo PCR product. Nanodrop reads 18.9ng/ul with an abnormally high 260/280. Could be impurities/salts from the gel.

Ligation of LovTwo into pGEM

18ul total volume

  • 8.5ul ligase buffer 2x
  • 1ul T4 ligase
  • 0.5ul pGEM vector
  • 8ul insert (Lovtwo)
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