Team:Northwestern/07 12

Notebook

Tuesday, July 12th

Agenda:

  • Transform pSB1C3 & miniprep
  • PCR cleanup/Gel

Tasks:

Jordan

  • Talked about ethics with CTD kids
  • PCR cleanup: used nuclease-free water instead of elution buffer, final conc. ~55 ng/uL
  • Added lab notes to Experiment

Michelle

  • Made 1L of 1X TAE buffer:
    • 100 mL 10X TAE
    • Filled to 1L dH2O
  • Made 100mL of 1% agarose:
    • 1 g agarose
    • 100 mL of 1X TAE
  • Gel prep
    • 3 uL SybrGreen
    • Allowed the gel to cool for a long time (~1.5 hours)
    • Used 5 uL of PCR product from linearizing the backbone and 1 uL purple loading dye
    • The agarose was still unusually soft
  • Website research

Paul

  • Outreach: Genedrive presentation
  • Transformed pSB1C3+Tet
  • PCR cleanup of linearized Tet backbone for Cas9 insertions

Sam

  • Did the outreach + made the outreach doc
  • Researched S. aureus gRNA
  • Responded to Rose
  • Refined CSS talents

Sara

  • Outreach at Roycemore
  • Emailed Bernd asking for empty backbones for saCas9 and spCas9

Shu

  • Updated our logo + graphics design research

Tasfia

  • Wrote/started lab notes on Experiment
  • Read up on mechanism of CRISPR/Cas9
  • Started a spreadsheet for iGEM parts we’re using

Tyler

  • Gel prep
  • Read up on saCas9 on protein structure/sgRNA design
  • Began designing gRNA gblock

Results:

Paul’s Tet-plasmid linearization worked, the others did not