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