Team:Northwestern/07 22

Notebook

Friday, July 22nd

Tasks:

Jordan

  • Met with Josh
  • Calculated transformation efficiency
  • Finished CLP letter, pending review
  • Emailed iGEM alums

Michelle

  • Made primers for extracting SS from GFP parts
  • Started inventory on the freezer

Sam

  • Met with Josh
  • Emailed Prof. DeLisa
  • Drafted an email to Eligo Bioscience

Sara

  • Miniprepped the 6 Cas9 Gibson cultures
  • Ran the restriction digest of the cas9 Gibson product again

Tasfia

  • Ran restriction digest of Gibson assembly Cas9(1)/Cas9(2)/Tet backbone with Sara
    • Recipe for 25-uL reaction
      • 0.6 uL undiluted (1730 ng/uL, so ~1ug) Gibson product
      • Diluted Gibson product from day before was ~43 ng/uL, but wasn’t used for digest because we had less than 20 uL of it remaining
    • 2.5 uL 10x buffer [CutSmart or NEBuffer4 (two digests were run with two different buffers)]
      • CutSmart 10x
      • NEBuffer4 10x
    • 1 uL EcoR1 restriction enzyme
    • 20.9 uL nuclease-free water
  • Ran two unsuccessful gels
    • First try: ladder bled through to the second (CutSmart) well
    • Second try: no DNA showed up (like yesterday), but ladder was fluorescent
  • Transformed sfGFP and mCherry plasmids with Shu
    • 50 uL cells in each CamR plate (one plate for each plasmid)
    • Followed bootcamp protocol with 50 uL cells and 450 uL SOC
  • Read some high-level antimicrobial resistance stuff to justify our project motivations for wiki

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