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
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
- Recipe for 25-uL reaction
- 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
