Method for constructing microsatellite multiplex PCR system of genome of Trionyx sinensis

Abstract

The invention discloses a technical method for rapidly constructing a multiplex PCR system of more than 10 * of Trionyx sinensis by using public data. the method is mainly characterized in that: 1) using a public database to obtain a genome sequence of Trionyx sinensis; 2) analyzing the genome sequence of Trionyx sinensis by using MISA software, designing primers in batches by using Primer 3, andcalculating the compatibility of polymorphic site primers by using AutoDimer software; 3) constructing a multiplex PCR system while reserving the 10-12 locus with the strong compatibility of primer sequence; according to the concentration of 0.4 mu M of upstream primer and 10 mu M of downstream primer of each locus, preparing the primer premix, and then preparing the working solution of multiplexPCR primer by mixing the premix of all loci in the same volume; and 4) amplifying that multiplex PCR system by a conventional PCR method and genotyping the amplification product by capillary electrophoresis. The multiplex PCR system constructed by the invention has the advantages of high mark flux, accurate genotyping result and high speed, and can provide a molecular tool for the research work ofthe genetics and breeding of the soft-shelled turtle of China and the like.

Description

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AI Technical Summary

Benefits of technology

This technology allows us to gather data on microscopic locations from an entire dataset quickly without requiring expensive equipment or materials that may be needed during testing. By doing this we save money while also providing precise measurements with faster processing times compared to other methods like fluorescent dye binding assays (FBA).

Problems solved by technology

This patented describes a new technique called microspike chromosomography (MC). It allows for quicker and easier manipulation of DNA fragments from various sources such as bacteria, yeast cells, moldings, plasma membranes, etc., making it ideal for studying these types of organisms. However, MC also faces challenges due to its slow process speed and expensive equipment required during preparative cloning processes. Therefore, the patents propose developing a faster multiplex PCR approach for identifying specific regions within each type of DNA segments.

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