A single primer capable of multi-species molecular labeling and its labeling method

Abstract

The invention relates to the field of biotechnology, especially to a single primer capable of multi-species molecular marking and a marking method thereof. The primer is designed according to the AT-rich intron region and regulatory region of a gene in a eukaryotic genome, and primer sequence length is 17 bp. A filling sequence is any sequence rich in GC bases; a core-region sequence is formed by random arrangement of AT bases; a selective base sequence is any base sequence. The primer can be used among eukaryote. Thus, synthesis cost of the primer is reduced, and utilization rate of the primer is greatly raised. Meanwhile, the DNA molecular marking method is based on a PCR reaction, polymorphism begins with intron sequence mutation and insertion / deletion. The method is simple and efficient; polymorphism is high; there are abundant amplified bands; results are reliable; and the bands are easy for separation and sequencing.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a single primer capable of multi-species molecular marking and a marking method thereof. 【Background technique】 [0002] DNA molecular markers are genetic markers based on nucleotide sequence variation among individuals. They can directly detect differences between biological individuals at the DNA level, and can directly reflect the genetic nature of organisms at the DNA level. They are genetic variations at the DNA level of individual organisms. a direct reflection of. It has the advantages of rich quantity, large amount of information, high polymorphism, detection is not affected by seasons, environments and individual developmental stages, simple and rapid detection methods, etc., and has been widely used in genetic diversity analysis, kinship evaluation, molecular fingerprinting, etc. Map construction, genetic linkage map construction, QTL or gene mapping and cloning, and mol...

AI Technical Summary

Problems solved by technology

Generally speaking, most of the DNA molecular labeling technologies that emerged later absorbed the advantages of the previous ones and overcome some of their shortcomings, so that they can be continuously optimized. Disadvantages such as small number, poor primer versatility, low primer annealing temperature, poor repeatability of primer amplification, and primers tend to amplify non-functional coding regions, etc.

[0004] The first inventor of the present invention applied for a Chinese patent in 2015, the patent number is: CN 104946742A, which discloses a single primer designed and synthesized for the gene exon region in the eukaryotic genome. The primer has a simple design , low synthesis cost, strong versatility, high utilization rate, high annealing temperature, good amplification repeatability, rich polymorphisms, etc. Design and synthesis of single primers containing subregions or regulatory regions, but there is no disclosure of the technology of using single primers designed based on gene intron regions or regulatory regions for multi-species molecular marker analysis

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