May 17, 2020
DH5α bacteria transformation protocol
- 1Ecole Normale Supérieure de Lyon
- CoudertLab
Protocol Citation: Yoan Coudert 2020. DH5α bacteria transformation protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bghijt4e
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 17, 2020
Last Modified: May 17, 2020
Protocol Integer ID: 37130
Thaw bacteria on ice
Dispense 50μl in a microtube
Add 1 to 5 μl of DNA, and carefully flick the tube to mix cells and DNA
Place the mixture on ice for 10 minutes
Heat shock at 42°C for 30 seconds
Place on ice for 5 minutes
Pipette 450 μl of room temperature SOC (or LB) into the mixture
Incubate at 37°C for 1 hour. Shake vigorously at 250 rpm.
Warm selection plates at 37°C
Spread desired volume (150-350 μl) onto selection plate
Incubate overnight at 37°C
Store at 4°C after incubation of needed