High-efficiency extraction method of peanut leaf DNA

Abstract

The invention belongs to the technical field of crops, and more specifically relates to a high-efficiency extraction method of peanut leaf DNA. The high-efficiency extraction method comprises following steps: peanut leaf samples are collected; CTAB solution is added into the collected peanut leaf samples in two steps; samples are smashed; the smashed peanut leaf samples are introduced into a waterbath pot for a certain time period of water bath; a chloroform/isoamyl alcohol solution is adopted for a first time of extraction; a phenol/chloroform/isoamyl alcohol solution is adopted for a secondtime of extraction; after extraction, an obtained supernate is collected, precipitation of DNA is carried out, and obtained DNA is washed and stored. The high-efficiency extraction method is capableof realizing high efficiency extraction of peanut leaf DNA at room temperature, and is especially suitable for extraction of DNA of large amounts of samples; extraction cost is relatively; extractiontime is short; batch extraction of a large amount of DNA can be realized; the quality of extracted DNA is basically the same with that of DNA extracted using DNA kits. Operation of the high-efficiencyextraction method is simple, and extraction cost is reduced by half or more of that of extraction using kits.

Description

technical field [0001] The invention belongs to the technical field of crops, and in particular relates to a method for efficiently extracting peanut leaf DNA. Background technique [0002] Peanut tissue DNA extraction is the first step in peanut genetic research and molecular biology experiments. The quality and efficiency of DNA extraction are directly related to the success or failure of subsequent experiments. Leaf tissue is the most commonly used material for DNA extraction. Phenolic compounds are one of the most diverse and widely distributed secondary metabolites in peanut leaves. These substances tend to interact with nucleic acids to form stable irreversible complexes when cells are broken. At present, there are two main techniques for extracting peanut tissue DNA: 1. Using biological kits, this method has high extraction quality, but the kits are expensive and the extraction cost is high; 2. Traditional CTAB method, this method has low cost, but the extraction Th...

AI Technical Summary

Problems solved by technology

At present, there are two main techniques for extracting peanut tissue DNA: 1. Using biological kits, this method has high extraction quality, but the kits are expensive and the extraction cost is high; 2. Traditional CTAB method, this method is low in cost, but the extraction The effect is not good and cannot meet the requirements of subsequent experiments

At the same time, the above two methods require the use of liquid nitrogen during the sample grinding process, resulting in slow grinding speed and low extraction efficiency, and the quality of DNA will be greatly reduced due to the oxidation of phenolic substances in the leaves.

Images