Team:Washington/Protocols/Transformation
From 2012.igem.org
General Transformation Protocol
- Thaw 20 μl of frozen competent cells on ice. Do not use glass tubes, which adsorb DNA.
- Add 20 μl DNA, pipette gently to mix (keep volume of DNA less than 5% of the cell volume)
- Incubate on ice for 30 minutes
- Note: If you are in a rush, you can shorten this incubation time to 5-10 min.
- Incubate cells for 45-90 seconds at 42°C.
- Incubate cells on ice for 2 min.
- Add 200mL of room temperature TB.
- Incubate for 1 hour at 37°C on shaker.
- Note: Can also save some time here by reducing incubation to ~45 min.
- Note: Step can be eliminated if plating on Amp plates, but not other antibiotics
- Spread 100-300 μl onto a plate made with appropriate antibiotic.
- Grow overnight at 37°C.
Adapted from [http://openwetware.org/wiki/Transforming_chemically_competent_cells_%28Inoue%29 OpenWetWare protocol].