Team:Northwestern/08 28

Notebook

Sunday, August 28th

Tasks:

Tasfia

  • Made 5 mL overnight cultures of "TetR-Cas9," "Assembled Cas9 from Cam Culture," GFP from iGEM kit
    • Three different colonies
    • Used 1000X Cam
    • Working concentration was 35 ug/mL
    • Added 5.15 uL to each culture tube
  • Made 5 mL overnight culture of gRNA Gibson product
    • One colony, duplicate overnight cultures
    • Used 1000X Tet (10 mg/mL)
    • Working concentration was 10 ug/mL
    • Added 5 uL to each culture tube
  • Gel extracted signal sequences (with homology added)
  • Used team-modified protocol:
    • Sat product for ~4 minutes between washes
    • Evaporated EtOH for ~12 minutes
    • Eluted in nuclease-free water after standing for ~15 minutes
  • Going forward: we can attempt a Cas9-SS Gibson after figuring out what’s wrong with the gRNA Gibson assembly

Tyler

  • Retransformed gRNA Gibson product from 08.26.16
    • 3 µL of gRNA Gibson assembly product
    • 2 µL positive Gibson kit control
    • 2 µL (-) control 1 µL transformation control
  • Retransformed 1 µL GFP from iGEM kit

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