Oct 06, 2021
Transformation
- 12021 iDEC NEFU_China
- NEFU_China 2021
Protocol Citation: Shuning Guo 2021. Transformation. protocols.io https://dx.doi.org/10.17504/protocols.io.bytupwnw
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: October 06, 2021
Last Modified: October 06, 2021
Protocol Integer ID: 53844
Abstract
This protocol is used to transform plasmid DNA into competent cells by chemical method or electroporation.
Materials
Equipment
MicroPulser Electroporator
NAME
Electroporator
TYPE
Bio-Rad laboratories
BRAND
1652100
SKU
Electroporation cuvette
Before start
Do not place the competent cells on the the ice for too long time before start.
Choose one transformation method depending on your requirement of transformation efficiency.
Chemical transformation8 steps
Thaw the competent cells on the ice for 10 min to let the suspension thaw.
Mix 100μl competent cells with about 10μl DNA.
Incubate on ice for ~30 min.
Heat shock at 42℃ for 45 seconds in the water bath.
Incubate on ice for 2~5 min.
Add 0.5~1ml LB medium into the mixture of cells and DNA and mix well.
Incubate at 37℃ for 1 hour, shaking at 200 rpm.
Plate on selective LB medium.