Transformation Protocol
New England Biolabs
Published: 2022-02-22 DOI: 10.17504/protocols.io.bd2yi8fw
Abstract
Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.
Before start
Steps
1.
Thaw competent cells On ice.
2.
Chill approximately 5ng (2µL) in a 1.5 ml microcentrifuge tube.
3.
Add 50µL to the DNA.
4.
Mix gently by pipetting up and down or flicking the tube 4–5 times to mix the cells and DNA. Do not vortex.
5.
Place the mixture On ice for 0h 30m 0s. Do not mix.
6. Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells’ manufacturer.
Heat shock at 42°C for 0h 0m 30s. Do not mix.
Note
7.
Add 950µL to the tube.
8.
Place tube at 37°C for 1h 0m 0s. Shake vigorously (250rpm,0h 0m 0s) or rotate.
9.
Warm selection plates to 37°C.
10.
Spread 50µL–100µL onto the plates.
11.
Incubate 1h 0m 0s at 37°C.
