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Team:Brown/Notebook/July5
From 2010.igem.org
Monday, July 5 2010
Transformation of XL1-B with ligations (failed)
Followed competent cell making protocol (quick CaCl2 method).
Had a small but visible pellet after centrifuging.
Started liquid culture of Gary’s XL1-Blues, added 1 mL of LB and incubated.
- 12 µl pGEM ligation
- 12 µl pNoTat ligation
- 5 µl RFP control DNA
Separate tubes, added our competent cells to each.
- Incubated for 20-30 minutes on ice (25 min).
- Heat shock 2 minutes at 42°C (actual: 44-45°C, don’t know if this affects anything.
- Plated 50 µl out on each amp++ plate
20 µl on each 1/3 of null plate.
Incubated at 37°C overnight; start at 7:15 PM.
