Extraction optimization method for chloroplast DNA from rhizoma polygonati

Abstract

The invention provides an extraction optimization method for chloroplast DNA from rhizoma polygonati. The extraction optimization method is characterized in that rhizoma polygonati leaves are used asmaterials, chloroplast is separated by a high-salt low-pH method, chloroplast genome DNA(cpDNA) is extracted by an SDS method, L9(3<3>) orthogonal experiments are performed, NaCl concentration in a buffer solution, leaf dosage and enucleation centrifugal force are used as inspection factors, and a rhizoma polygonati cpDNA extraction technology is optimized. The result indicates that when the NaClconcentration in the buffer solution is 1.50 mol/L, the leaf dosage is 10g, the enucleation centrifugal force is 600g, the extracted cpDNA does not have nuclear genome pollution and is good in qualityand high in production rate, and the condition that the NaCl concentration in the buffer solution is 1.50 mol/L, the leaf dosage is 10g and the enucleation centrifugal force is 600g is the optimum condition of the experiment. The method is simple, convenient and quick, is not high in requirements for experimental instruments, low in cost and high in yield, and can provide reference for extractionof the cpDNA from rhizoma polygonati plants.

Description

[0001] 1. Technical field [0002] The invention belongs to the technical field of molecular biology, and in particular relates to a method for extracting and optimizing Polygonatum chloroplast DNA. [0003] 2. Background technology [0004] Chloroplast is the key site for photosynthesis in green plants, and has relatively independent genetic material—namely, chloroplast genomic DNA (Chloroplast DNA, cpDNA). Most of the chloroplast genomes of higher plants are 120-160kb, and their DNA molecules are in the form of covalent, closed, circular double strands. The existence of the chloroplast genome was confirmed by Hans and Walter in 1962 \ . In 1986, the complete chloroplast genome sequence of tobacco (Nicotiana tabacum) and ground money (Marchantia polymorpha) was reported for the first time. In recent years, with the wide application of high-throughput sequencing technology in chloroplast genome sequencing, the number of chloroplast genome sequences However, due to the diffic...

AI Technical Summary

Problems solved by technology

[0006] Polysaccharides, polyphenols and other secondary substances in Polygonatum are high in content, which affec

Images