A large amount of oat seed dna extraction and purification method suitable for pcr amplification

Abstract

An oat seed DNA large-scale extraction and purification method suitable for PCR amplification is disclosed. According to the method, DNA is extracted directly from oat seeds; during an extraction process, a centrifuge tube having a volume of 50 mL is utilized to sample and extract the DNA; the concentration of a CTAB extraction buffer solution is 3% which is increased by 1% than that in conventional methods; a phenol-chloroform (V:V=1:1) solution and a chloroform-isoamylol (V:V=24:1) solution are separately used for extraction for one time during the extraction process; and the chloroform-isoamylol (V:V=24:1) solution is used for purification extraction for one time during a purification process. The high-quality high-purity oat genome DNA is successfully extracted. The period for seed germination to form plantlets is saved. The yield and the quality of the extracted DNA meets further PCR analysis. A DNA sample extracted by the method is applied for oat SSR molecular marking. The method lays a foundation for further oat molecular genetic research.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a large-scale extraction and purification method of oat seed DNA suitable for PCR amplification. Background technique [0002] Oat (Avena L.) is one of the most important crops in the world. It ranks sixth in the planting area and total output of cereal crops in the world. It has high nutritional, edible, health care and feeding values. So far, there are more than 3,000 oat resources in my country's crop germplasm resource bank. It is an urgent subject to carry out in-depth research on these materials by using modern molecular biotechnology. [0003] With the in-depth development of modern molecular biology technology in the field of crop research, PCR-based molecular marker technology has been widely used in the fields of seed purity identification, seed transgenic detection, and germplasm resource diversity research, such as RAPD, AFLP, SSR, etc. , ISSR, SRAP and SNP e...

AI Technical Summary

Problems solved by technology

[0004] The extraction methods of oat DNA mainly include SDS, CTAB method and kit method. The kit method is expensive and not suitable for identification and analysis of a large number of samples. The SDS method can only extract a small amount of DNA samples, which cannot meet the requirements of modern molecular biology. A large number of tissue DNA analysis requirements in research, and the effect of CTAB method on oat DNA extraction is better than SDS, therefore, it is necessary to use a large number of CTAB method to extract genomic total DNA when doing molecular markers such as AFLP, SSR and ISSR

Although the DNA extraction method has been optimized by many researchers, most of the materials are collected from the leaves, and the oats need to grow to a certain stage when collecting materials, which takes a long time

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