Transform PCR and test plasmids into competent cells
- Four Samples for transformation
- Novagen Test Plasmid into NovaBlue GigaSingles (recently purchased)
- Novagen Test Plasmid into M15MA competent cells (recently made) - cells sent from David Tirrell at Caltech
- A71M M8S M33S M103S Ascaris Hb into NovaBlue GigaSingles (recently purchased)
- WT Ascaris Hb into NovaBlue GigaSingles (recently purchased)
- 1.875 g LB
- 1.5 g Agar
- 75mL water
- 75uL of 100mg/mL ampicillin after mixture had cooled but not set
- Place plastic culture tubes on ice for 15 minutes
- Place DNA for transformation on ice
- After DNA has thawed, place cells on ice. (Note - want to minimize the time that the cells are out of freezer and on ice)
- Add 1uL of DNA to the appropriately labeled culture tube and let sit in ice for 1 minute
- Add cells (30uL, 40uL, or 50uL) to the tube on top of the DNA and gently pipette up and down to mix cells and DNA
- Put tube back in ice for 4 minutes
- Heat shock at 42C for 80seconds
- Add 100uL of SOC media
- Shake at 37C for 1 hour
- Plate 50uL of culture media
- Add data and results here...
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