Transformation
Ana Belem García González, Georgina Diego, Jair Alexis Gardea Sáenz, Irán Alessandra Chaparro Rodríguez
Published: 2022-10-09 DOI: 10.17504/protocols.io.36wgq7xpyvk5/v1
Abstract
A protocol for transforming E. coli cells is shown.
Steps
Transformation
1.
Mix 50µLof fresh competent cells with2µLof genetic material (softly without pipetting) in a sterile 1.5 mL Eppendorf tube.
2.
Incubate On ice for 0h 30m 0s
3.
Apply thermal shock in a water bath at 42°Cfor 0h 0m 30s
4.
Immediately, put On ice
5.
Add250µL of SOC medium (tempered at 37°C).
6.
Incubate at 37°C for1h 0m 0s at 225rpm.
7.
Disseminate 50µLin a selective medium and incubate 1h 0m 0s at 37°C
