Prokaryote molecule label method based on RNA polymerase binding site

Abstract

The present invention discloses a prokaryote molecule label method based on a RNA polymerase binding site. According to the method, primers are designed and synthesized according to the prokaryote RNA polymerase binding site; the genomic DNA of the prokaryote is extracted as a template; the designed primers and the extracted DNA are adopted to carry out touchdown PCR amplification; the amplified product is separated by gel electrophoresis to detect the difference of the spacer region of the prokaryote RNA polymerase binding site. The molecule label method of the present invention has characteristics of high specificity, abundant labels, convenient operation, high stability, good repeatability, and easy separation and sequencing of the band, and can be used in the prokaryote researches, wherein the researches comprise heredity, germplasm identification, diversity analysis, genomics, assisted breeding and the like.

Description

Technical field [0001] The invention belongs to the field of biotechnology and relates to a prokaryotic biomolecule marking method. Background technique [0002] DNA molecular markers are biological characteristics that can clearly reflect genetic polymorphism, and are a direct reflection of the variation of organisms at the DNA level. It has the advantages of large quantity, rich information, high polymorphism, has nothing to do with gene expression, and is not affected by the environment and individual development process. It has been widely used in breeding, resource and gender identification, genetic relationship analysis, genetic diversity evaluation, Genetic mapping, gene mapping and cloning, etc. [0003] Since Botstein et al. first proposed RFLP (Restriction Fragment Length Polymorphism) markers in 1980, more than a dozen molecular markers have been developed. RFLP is the first-generation molecular marker based on molecular hybridization. There are many early applications...

AI Technical Summary

Problems solved by technology

The advantages of these two markers are simple operation, medium yield, and easy separation and sequencing of bands, but there is also a common defect: the initial annealing temperature is low, and false positives may occur

[0004] It can be seen from the above that since the 1990s, molecular labeling technology has continued to develop and new labels have emerged, but existing labels still have some defects. Therefore, it is necessary to develop new labeling methods that are simple, fast, and highly stable very necessary

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