Team:Cornell/Notebook/Salicylate reporter

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June

Week

June 29th, Friday

  • Vent PCR at 11:00 (DPW)
    • Dylan's magic triple anneal method (55/60/63)
  • Gel purified PCR product (DPW)
    • Quantified product at 22.4 ng/uL
    • Set up digestion of p21 PCR product with EcoRI-HF and AscI.
      • 22 ng/uL --> 45.5 uL sample for 1 ug digest
      • Buffer 4
    • Ran digestion on gel.
    • Sliced out relevant band on gel, stored overnight at -20.
  • Miniprepped overnight cultures of PL14-PL20 (STC)
    • Using C1015 rotor, 6666 x g, the Corning culture tubes only fit in the centrifuge with the lids off
  • Made LB, 3x 60 mL in 100 mL bottles (SS)
  • Made LB Agar, 4 x 250 mL LB Agar in 500 mL bottles (CS)
  • Autoclaved LB, LB Agar, and milliQ (SS)
  • Made LB plates with Kan (CMR)
  • CUGEM movie outing at 8:00 pm.
  • Phusion PCR at 10:00 pm (DPW)
    • Dylan's magic triple anneal method (57/65/70, 35 cycles total)
    • Amplifying nah operon from Gibson 1
    • Appending BioBrick cutsites for ligation into pSB3C5

June 30th, Saturday

  • Took PCR out of thermal cycler at 9:00 am (DPW)
    • Set of gel using NEB 100bp and 2-log ladders (10:00am)
  • Continued gel extraction of p21 PCR digest from previous day (SS)
  • Set up ligation of p21 PCR digest extraction and dephosphorylated pBBRBB+mtrB (SS)