Oct 24, 2020
Transformation
- 12020 iGEM NEFU China
- 2020 iGEM NEFU China
Protocol Citation: Zhujun Wei 2020. Transformation. protocols.io https://dx.doi.org/10.17504/protocols.io.bnwwmffe
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 24, 2020
Last Modified: October 24, 2020
Protocol Integer ID: 43702
Materials
DH5α chemically competent
LB medium
Safety warnings
Please wear gloves for the experiment.
Put DH5α chemically competent bacterium on ice for melting about 15min.
Add 50 μl DH5α chemically competent bacterium into 1.5 ml tubes, mix with ligation product or digested product thoroughly, incubate the bacterium on ice for 30 min.
Heat shock the bacterium for exactly 45 s at 42 ℃ water bath.
Place back on ice for 2 min.
Add 500 μ l LB medium to each tube, shake the tubes at 37 ℃, 200 rpm for 45 min.
Centrifuge the bacterium at 3000r for 3 min.
Discard 300 μ l supernatant and resuspend the sediment.
Plate 100 μl for a reaction product.
Incubate plates upside down overnight at 37℃.