Pumpkin SSR labeled primers applied to multiplex PCR reaction and application of pumpkin SSR labeled primers

Abstract

The invention discloses three groups of pumpkin SSR labeled primers applied to multiplex PCR reaction, belongs to the technical field of plant molecular biology DNA marks, and discloses an application of the primers to identification of pumpkin varieties and a pumpkin variety identification method. A batch of pumpkin SSR marks are developed by using pumpkin transcriptome sequencing results; three marks are selected from the marks and are characterized in that the polymorphism is high, the strip pattern is excellent, the size difference of amplification products is about 50bp, and primers cannot be matched with each other for forming hairpin structure SSR loca; by virtue of optimization of a PCR reaction system and an amplification program, a multiplex PCR analysis technical system with three SSR loca is built; the SSR marks are excellent in multiplex PCR amplification strip pattern, high in polymorphism and easily readable in results; the analysis efficiency of SSR is remarkably improved; the SSR marks can be applied to the fields of identification of pumpkin varieties, construction of fingerprint maps and identification of purity of hybrid seeds.

Description

technical field [0001] The invention belongs to the technical field of molecular biology DNA markers, and in particular relates to a set of pumpkin SSR marker primers suitable for multiple PCR reactions, and also relates to the application of the primers in pumpkin variety identification and a pumpkin variety identification method. Background technique [0002] Simple sequence repeat (simple sequence repeat, SSR) refers to the 1-6bp base tandem repeat sequence existing on the genome, which shows polymorphism among different individuals due to the difference in the number of repeats. Simple repeats have been used extensively to develop SSR markers. Due to the advantages of high polymorphism, good repeatability, co-dominant inheritance and gene coverage, SSR markers are widely used in the fields of genetic map construction, gene positioning and variety identification. [0003] The SSR marker is a sequence-specific marker. To develop an SSR marker, you must know the SSR site a...

AI Technical Summary

Problems solved by technology

However, due to the existence of unfavorable factors such as the interference of different primer pairs in the same PCR reaction, which leads to the failure of PCR amplification, and the difficulty in reading the results due to the increase in the number of amplified products, it is very difficult to establish a multiplex PCR technology for SSR markers.

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