Plasmids are a class of double-stranded, closed-loop DNA ranging in size from 1 kb to over 200 kb . In general, the plasmid can be stably and stably in an extrachromosomal free state and has an autonomous replication function; but under certain conditions, it can also reversibly integrate into the host chromosome, replicate with the replication of the chromosome, and be transmitted through cell division. Descendants. Plasmids have become the most commonly used vector for nucleic acid cloning and sequencing. The yield and purity of plasmid DNA extraction is directly related to the success or failure of downstream experimental procedures. There are many methods for extracting plasmid DNA . The most widely used method is the alkaline lysis-neutralization method. The basic principle is: when the bacteria are lysed in NaOH and SDS solution, the protein and DNA are denatured; when the neutralizing solution is added, The plasmid DNA molecule can be rapidly renatured and dissolved, and remains in the supernatant during centrifugation; the protein and chromosomal DNA can not be renatured and flocculent, and can be precipitated during centrifugation. Plasmid DNA purification process typically utilizes a relatively small plasmid DNA and covalently closed both properties, e.g., cesium chloride - ethidium bromide equilibrium gradient centrifugation, ion exchange chromatography, gel filtration chromatography, polyethylene A method such as fractional precipitation of alcohol. But these methods are relatively expensive or time consuming. For small-volume plasmid DNA , residual proteins and RNA can be removed by simple steps such as phenol, chloroform extraction, RNase digestion and ethanol precipitation . The purified plasmid DNA can be used for enzyme digestion, transformation, probe labeling, sequencing, etc. Claim. However, this method takes a long time and requires contact with toxic solvents such as phenol and chloroform, and the extracted plasmid DNA is sometimes not completely digested by the restriction endonuclease. Therefore, in the pursuit of efficiency, this method has been gradually obtained with high quality. The quick and convenient silica gel membrane centrifugal adsorption column purification method is replaced.
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