Oct 24, 2020
Dot mutation
- 12020 iGEM NEFU China
- 2020 iGEM NEFU China
Protocol Citation: Zhujun Wei 2020. Dot mutation. protocols.io https://dx.doi.org/10.17504/protocols.io.bnwxmffn
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: 43703
Safety warnings
Please wear gloves for the experiment.
The entire plasmid was amplified reversely by PCR using primers with the fragment sequence that you want to replace.
The temple plasmids in the PCR process were digested by DpnI enzyme.
The digested product was transferred into DH5α. Overnight culture them at 37℃.
To determine whether the vector was constructed successfully, colony PCR and enzyme digestion were done.
Select the positive results for sequencing and the final results were obtained.