Team:Georgia State/Notebook

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Contents

Notebook

July 14, 2010

Virginia, Joe, Angie, Alykhan

  • 10 glycerol stocks of 12E
  • P. pasoris competent cells
  • Started, ready at 2pm
  • Plamid resuspension buffer made
  • LB agar aliquots
    • warm in water bath
    • one 10mL tube for 1L agar
  • extraction of 12E+9A plasmid parts: white tubes in freezer

July 13, 2010

Virginia

  • 10 glycerol stocks of each
    • P. pastoris
    • 9A E. coli
  • Inoculated 12E broth for plasmid extract and glycerol stocks
  • Plate of P. pastoris for quality control

July 8, 2010

Angie, Kendra, Melissa, Nishedh, Alykhan

  • Diluted pichia cells in a volume of 50mL to a OD600 = .186+.201
  • Transform pars 12E and 12M into E. coli

July 7, 2010

Joe, Kendra, Angie

  • Replated colonies from 9A RFP plate (3 plates).
  • Colonies on broth 100 µL for 9A but no fluorescence observed
  • 9A survivor colonies replated onto 10 µg/mL ampicillin plates and incubated at 30°C
  • Mother plate in fridge
  • Made 1000mL YPD
  • Growing P. anomala in YPD broth for competent cells protocol

July 6, 2010

Dan

  • Replated E. coli culture

Alykhan

  • Transformation 9A, 8I (digested) & GFP.

July 2, 2010

Alykhan and Virginia

  • DNA purification and ligation
  • Replated E. coli culture

July 1, 2010

Joe

  • Plated transformed cells containing either GFP (control) or iGEM part onto LB plates with 10µg/mL ampicillin (2 plates each, 10mL or 100mL)
  • Also plated ≈100mL (remaining amount) of cells onto LB plates not containing ampicillin.
  • Spread plates with hockey stick and placed in 37°C at 7:35.