Extraction of plasmid DNA from Escherichia coli (alkaline lysis)

Summary

The basic principle of plasmid DNA extraction by alkaline lysis is that solution I suspends the cells and EDTA can chelate with Ca2+ and Mg2+ to inhibit the activity of DNase, solution II lyses the bacteria, denaturing proteins and a small amount of plasmid, and solution III causes most of the proteins to co-precipitate with the bacterial genomic DNA, and the plasmid DNA is denatured. This method is suitable for the extraction of small amount of plasmid DNA, and the extracted plasmid DNA can be directly used for enzymatic PCR amplification.

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