Method for screening microorganism high-polymorphism molecular marker sites

Abstract

The invention discloses a method for screening microorganism high-polymorphism molecular marker sites. The method comprises the following steps of uniformly mixing microspecies of different organismsat equal amount, and extracting total nucleic acid; building a high-flux sequencing library; finding variation sites on a genome; screening the high-polymorphism sites in a gliding translation way; designing multiple amplification primers at both sides of a candidate site; screening the multiple primers, so as to obtain the novel high-polymorphism molecular marker sites. The method has the advantages that in theory, all available high-polymorphism molecular marker sites on the genome can be screened at one time, and be directly used for high-flux detection of variety, and the repeated demonstration process in the traditional screening method is not needed; the screened molecular marker sites can be detected in batch, and the respective molecule amplification and detection on each molecularmarker site are not needed, so that the detection speed is accelerated, and the accuracy is improved; the screened molecular marker has high polymorphism and good resolution, the screening process issimple and quick, and the process is standard.

Description

technical field [0001] The invention discloses a method for screening microbial high polymorphic molecular marker sites, belonging to the field of biotechnology. Background technique [0002] Molecular markers refer to specific DNA fragments that can reflect certain differences in the genomes of organisms or populations. Molecular marker technology is widely used in fields such as gene positioning, genetic map analysis, forensic medicine, and approval of new varieties. It has extensive application value and significance of theoretical research. However, due to the limitations of existing molecular marker development methods, the number of available molecular markers is very small, and even some species lack available molecular marker sites. In the process of realizing the present invention, the inventors found at least the following problems in the prior art: 16S rDNA can only be distinguished into species, but cannot be distinguished at the race level; the development of t...

AI Technical Summary

Problems solved by technology

However, due to the limitation of the development methods of existing molecular markers, the number of molecular markers currently available is very small, and even some species lack available molecular marker sites

In the process of realizing the present invention, the inventors found at least the following problems in the prior art: 16S rDNA can only be distinguished into species, but cannot be distinguished at the race level; the development of traditional high polymorphic molecular markers mostly relies on The researcher's personal experience is obtained through a large number of verification experiments. The process is costly and time-consuming, and it may not be possible to obtain truly high polymorphic marker sites

Moreover, the entire development process can only find out one highly polymorphic molecular marker site at a time, but there is nothing to do with the many potential high polymorphic molecular marker sites existing on the genome

Traditional high-polymorphic molecular markers are mostly used for the detection of one marker and one marker, and cannot detect multiple molecular marker sites in a large-scale and high-throughput manner.

Some species currently have abundant variation site information, but it may not be applicable to all species

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