High-flux quick extraction method of single rapeseed genome DNA (Deoxyribonucleic Acid)

Abstract

The invention provides a high-flux quick extraction method of single rapeseed genome DNA (Deoxyribonucleic Acid), belonging to the biotechnology. The method comprises the following steps of: sprouting, taking the material, splitting, extracting, precipitating, centrifuging, drying, dissolving and preserving, replacing a water bath by a PCR (Polymerase Chain Reaction) instrument, and using a 96-pore PCR plate in cooperation with a multi-channel pipettor to operate in the whole process of the extraction. The method has the characteristics that the time for extracting the rapeseed genome DNA is short, the cost is low, the operation is convenient, the flux is high, the subsequent DNA absorption is convenient, and the output and the quality of the extracted DNA completely meet the need of PCR reaction.

Description

technical field [0001] The invention relates to a method for extracting genome DNA of rapeseed seeds, which belongs to the field of biotechnology. Background technique [0002] With the in-depth development of modern molecular biology research, various biotechnology methods are becoming more and more mature, especially PCR-based molecular marker technology, which is widely used in the fields of seed purity identification, seed transgenic detection, and germplasm resource diversity research. DNA is the carrier of biological genetic information. Whether it is molecular marker technology or genetic transformation and identification, the premise is the extraction of genomic DNA. [0003] At present, the methods used for rapeseed DNA extraction include SDS method, CTAB method, high-salt method and kit method. The extraction methods have been optimized by many researchers, but most of them are carried out for rapeseed leaves. The process takes a long time, so the method of ex...

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Problems solved by technology

Zhang Fuli et al. (Agricultural Science and Technology (English Edition), 2012, Volume 13, No. 3, 485-488) mixed multiple seeds, ground them into powder, took 100 mg of powder and added high-salt extract to extract genomic DNA. This method Although rape seedling growth and liquid nitrogen grinding are avoided, toxic organic solvents such as phenol and chloroform are used, and the steps are still relatively cumbersome; Xie Jingmei et al. Genomic DNA was extracted from germinated seeds, and the improved CTAB method was used, and the extraction effect was better. However, this method still needs steps such as liquid nitrogen grinding and chloroform extraction, and has not been greatly improved.

[0004] In summary, the current methods for extracting rapeseed genomic DNA generally have disadvantages such as cumbersome operation, long extraction time, and high cost.

In addition, there are few reports on genomic DNA extraction using rapeseed seeds as materials, but in order to ensure the purity of rapeseed hybrids, hybrid purity identification must be carried out within a limited time from field harvest to packaging, and rapid extraction of rapeseed seeds is required. Genomic DNA, previous DNA extraction methods were not adapted to this need

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