Electroporation
Oct 18, 2020
Open access
Protocol CitationJiaxin Li 2020. Electroporation. protocols.io https://dx.doi.org/10.17504/protocols.io.bni7mchn
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: Oct 18, 2020
Last Modified: Oct 18, 2020
PROTOCOL integer ID: 43327

Public workspaceElectroporation

  • 1South China University of Technology
1
Separate the bacterial liquid in shaking tube into 1.5 mL (2 tubes) per tube, centrifuge quickly for 90 s at 4℃ and 12000 rpm.
2
Add 1 mL of 300 mM sucrose solution to each tube of bacterial liquid, centrifuge for 2 min at 12000 rpm and 4℃, and pour it out.
3
Resuspend the competent cells (tube one) by adding 100 μl 300 mM sucrose solution, then transfer the liquid to tube two to resuspend the competent cells, and divide into 50 μl tubes (two tubes in total).
4
Add 1 μg plasmid into each tube, gently blow and mix.
5
Add 50 μl of suspension to the electroporation cuvettes.
Electroporation
6
Turn on electroporator and set to 3kv, 400 ohms and 25 µF
7
Take out the liquid culture medium of LB with 37℃ heat preservation for standby.
8
Transfer the DNA-cell mixture to the cold cuvette, tap on countertop, wipe water from exterior of cuvette and place in the electroporation module and press pulse.
9
Immediately add LB culture solution, mix by pipetting up and down once and transfer to a microcentrifuge tube, 5 ml culture tube.
10
Place in the shaker/incubator at 37°C incubator for 1 h-2 h
11
Incubate overnight at 37°C