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Optimization of mass extraction conditions of plasmid DNA in Escherichia coli

Date:2021/01/06 13:53:59
The principle of alkaline lysis method was used to extract a large amount of plasmid DNA from E. coli, and the relevant steps of the plasmid extraction kit were optimized. The results show that the plasmid DNA extracted by this method according to the optimized steps has high concentration and good quality, which can meet the requirements of routine experiments such as subsequent enzyme digestion.
Plasmids are closed circular DNA molecules that exist on the chromosomes of bacteria, yeasts, and actinomycetes and express the genetic information they carry. They are often used as vectors in genetic engineering. Insert the target gene fragment into the plasmid to form a recombinant plasmid or recombinant body. Then, the reconstituted recombinant plasmid is treated and transformed by chemical reagent methods such as electric shock method and CaCl2, and then transferred to the recipient bacteria (such as Escherichia coli) to make the target gene Reproduce or express in the recipient bacteria. By optimizing the steps in the kit extraction of plasmids, after DNA concentration measurement, it was found that the quality and purity of the plasmid DNA extracted by the optimized steps were better than those of the control group, and could better meet the requirements of subsequent experiments.
1Materials and methods

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