Team:Northwestern/08 31

Notebook

Wednesday, August 31st

Tasks:

Michelle

  • Miniprepped gRNA overnight colony
    • 216.0 ng/uL, 260/280: 1.87, 260/230: 2.20
  • Made nfH2O aliquots
  • Autoclaved things
  • Grew up TetR-Cas9 for western blot
    • 50 mL culture
    • Starting OD600: 0.035 A
    • 1 hour OD600: 0.048 A
    • 2 hour OD600: 0.157 A
    • 2.5 hour OD600: 0.249 A
    • 3 hour OD600: 0.464 A
    • 3.25 hr OD600: 0.600 A

Tasfia

  • Gel extracted PCR products (stored in -20°C and inputted in “Freezer (-20°C) Inventory”) with Leonard lab IBI gel extraction kit, using all iGEM modifications
    • sfGFP/mCherry for GG
    • Tet-Lrz-GFP
    • Cas-Lrz-SS

Tyler

  • Gibson Reaction for Ycdo into Lnrzed Cas9
    • 1.56 µL backbone
    • 0.29µL insert (Ycdo)
    • 3.15µL water
    • 5 µL Gibson mix
  • Negative Control
    • 1.56 µL
    • 3.44 µL water
    • 5 µL Gibson mix
  • Positive Control
    • 5µL (+) control
    • 5µL master mix
  • Transformation
    • 1 µL (+) transformation control (CamR)
    • 2 µL Gibson (+) control (AmpR)
    • 2 µL Gibson (-) Control (CamR)
    • 3 µL Gibson Product (CamR)
    • 3 µL gRNA Product (TetR)

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