Transformation using electroporation
Ashwinuday
Published: 2021-09-12 DOI: 10.17504/protocols.io.bx6fprbn
Abstract
For transforming cells with the DNA of interest using electroporation as the tool.
Steps
1.
To an electroporation cuvette add50µL electro-competent cells of the required strain, 70-100ng of DNA of interest and 50µL of autoclaved ultrapure water (milliQ)
2.
Keep the cuvette in ice for 0h 20m 0s
3. Ensure that there is no sparking
Pulse the cuvette using a pulser using the bacteria setting at 2.5keV
Safety information
4.
To the cuvette add 2X LB up to same volume
5.
Incubate the cuvette at37°C for 0h 30m 0s
6.
Spread the contents of the cuvette onto an LA plate with appropriate antibiotics
7.
Incubate the plate overnight at 37°C
