Sep 12, 2021

Public workspaceTransformation using electroporation

DOI
dx.doi.org/10.17504/protocols.io.bx6fprbn
  • 1Indian Institute of Science Education and Research, Pune
  • iGEM IISER Pune India 2021
Ashwinuday
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DOI: dx.doi.org/10.17504/protocols.io.bx6fprbn
Protocol Citation: Ashwinuday 2021. Transformation using electroporation . protocols.io https://dx.doi.org/10.17504/protocols.io.bx6fprbn
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: In development
We are still developing and optimizing this protocol
Created: September 12, 2021
Last Modified: September 12, 2021
Protocol Integer ID: 53159
Abstract
For transforming cells with the DNA of interest using electroporation as the tool.
Safety warnings
The electro-competent cells should be done thoroughly to ensure complete washing away of extra ions which might cause sparking while pulsing
To an electroporation cuvette addAmount50 µL electro-competent cells of the required strain, Amount70-100 ng of DNA of interest and Amount50 µL of autoclaved ultrapure water (milliQ)


Keep the cuvette in ice for Duration00:20:00

20m
Pulse the cuvette using a pulser using the bacteria setting at 2.5keV

Safety information
Ensure that there is no sparking

To the cuvette add 2X LB up to same volume
Incubate the cuvette atTemperature37 °C for Duration00:30:00

30m
Spread the contents of the cuvette onto an LA plate with appropriate antibiotics
Incubate the plate overnight at Temperature37 °C