Method for extensively screening scallop SNP

Abstract

The invention belongs to a method for extensively screening scallop SNP in the technical field of the scallop DNA molecular genetic marker, which comprises the following steps that: solution D and phenol-chloroform are firstly used for extracting a plurality of Chlamys farreri RNA, and a ultraviolet specrophotometer is used for measuring the concentration of the mixture of solution D and the phenol-chloroform which are uniformly mixed; a scallop full-length cDNA sample is re-established and comprises the synthesis of a cDNA first chain and the synthesis and augmentation of a cDNA second chain; then homogenization of full-length cDNA is performed, and ultrasonic breaking is performed; then sequence measuring joints are connected, and terminal repair, joint connection and sample augmentation are included; finally a biological software is used for analyze the data to obtain an SNP marker; then an SNP marker to be screened is selected to design a primer; and DNA of individual scallop is extracted to be equivalently mixed to be used as a template, conventional colony sequence measuring is undertaken after the PCR augmentation, and the position point with the occurring frequency of single base difference being 10 percent or more is defined as an SNP marker. The method has safe and reliable principle, simple and controllable flow procedures, reliable running of large-scale screening, excellent effect and strong practicability.

Description

Technical field: [0001] The invention belongs to the technical field of scallop DNA molecular genetic markers, and relates to a method for large-scale screening of scallop SNP markers. Background technique: [0002] Due to their large number and stable heritability, SNP markers are widely used in the construction of high-density genetic linkage maps and the QTL mapping of trait-related functional genes. As one of the most important marine economically cultured shellfish in my country, scallop occupies an extremely important position in the marine aquaculture industry. Large-scale screening of scallop SNP markers is crucial for the construction of high-density genetic maps of scallops, the fine mapping of QTLs related to economic traits, and further molecular design breeding. At present, large-scale screening of SNP markers is widely used in humans and model organisms with rich genomic information. However, no efficient and low-cost large-scale screening methods have been re...

AI Technical Summary

Problems solved by technology

However, due to the large genome of scallop with 1.2G and many repetitive sequences, the separation efficiency of whole genome sequencing is relatively low. Analysis software is used to screen SNP markers. On the one hand, it avoids repetitive sequences and improves screening efficiency; on the other hand, it can simultaneously obtain SNP markers directly related to functional genes, which has the characteristics of high throughput, high efficiency, and low cost. An effective method for screening scallop SNP markers is becoming a subject of research and development by a large number of researchers in this technical field