Team:Northwestern/08 11

Notebook

Thursday, August 11th

Tasks:

Paul

  • Submitted sequencing of the 08.09 miniprep (w/ antibiotic) of Jordan’s 1:3 Gibson
    • “1”= VF’
    • “2”= Cas9_Seq1
    • “3”= Cas9_Seq2
    • “4”= Cas9_Seq3
  • PCR of Tet lnrz for GFP (DpnI for 2 hr @ 37°C)
  • Presentation for profs

Sam

  • Provided Building with Biology to UNSW (and verified that you’re allowed to do that)
  • Told them about our experiences so far
  • Updated outreach log
  • Improved Collaboration slide in presentation
  • Updated Jamboree costs spreadsheet
  • Additional Eligo editing
  • Autoclaved Paul’s NaCl the normal way (and also some nearby diH2O just for kicks)
  • Found out that autoclaving kills DNases but not RNases

Sara

  • Made GG PCR parts fmol dilutions

Tyler

  • Made Sequencing premix rxns for Jordan's 1:3 Cas9 08.09.16 miniprep
    • 3.7 µL of 1:3 Jordan Cas9
    • 1.2 µL of each primer (VF, Seq1, Seq2, Seq3)
    • 10.1 µL of water
  • RTW PCR of Cas9 part for insertion of signal sequences
    • 2x 50 µL tubes (one with DMSO, one without)
      • 21.5 µL nf water/22.5 µL nf water (tube w/o DMSO)
      • 1 µL DMSO/0 µL DMSO
      • 0.5 µL Cas9 Template (Jordan’s 1:3)
      • 1 µL SS_Lnrz_FWD
      • 1µL SS_Lnrz_RE
      • 25 µL Taq Master Mix
    • 1x50µL negative control
      • 22 µL nf water
      • 1 µL DMSO
      • 1 µL SS_Lnrz_FWD
      • 1 µL SS_Lnrz_REV
      • 25 µL Taq Master Mix
    • DpnI digest for 2 hours at 37°C (0.5 uL/tube)
  • Presentation for profs

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