PCR detection method for identifying germplasms of four populations of Pelodiscus sinensis, primer group and kit

Abstract

The invention discloses a PCR detection method for identifying germplasms of four populations of Pelodiscus sinensis, a primer group and a kit. The primer group comprises primers of SEQ No.1 / SEQ No.4, SEQ No.2 / SEQ No.5 and SEQ No.3 / SEQ No.6. The kit comprises a 2*TaqPCR premixed liquid, a 5U / mul enzyme BglI and 10*digestion buffer liquid A, a 5U / mul HpaII and 10*digestion buffer liquid B, a 5U / mul ClaI and 10*digestion buffer liquid C, 10mg / ml BSA, a contrast DNA template and a 10pM amplimer. The method comprises the following steps: carrying out PCR amplification of extracted DNA through using the primer group, digesting to obtain a digestion product, carrying out agarose gel electrophoresis, and comparing the obtained electrophoresis result with a standard electrophoretogram to obtain a result. The detection method has the advantages of simplicity, good reaction stability and repeatability, and cost saving.

Description

technical field [0001] The invention belongs to the field of aquatic germplasm detection, and relates to a PCR detection method for germplasm identification of four populations of soft-shelled soft-shelled turtle, a primer set and a kit, and specifically relates to a germplasm identification of new species of Chinese soft-shelled turtle Taihu population, Taiwan population, Yellow River population and Japanese strains of Chinese soft-shelled turtle The rapid detection method and its primer set and kit. Background technique [0002] Chinese soft-shelled turtle ( Pelodiscus sinensis ), commonly known as turtle, soft-shelled turtle, etc., belongs to the genus Turtleidae of the order Turtles of the order Reptiles, and is widely distributed throughout my country. It has now developed into an important famous and special aquaculture species in my country. According to statistics, in 2012, the national soft-shelled turtle breeding output reached more than 330,000 tons, with an outp...

AI Technical Summary

Problems solved by technology

[0003] According to the requirements of the current Chinese soft-shelled turtle germplasm standard GB21044-2007, the external morphology, internal structure, biochemical isozymes, chromosome karyotype and other methods are mainly used for germplasm identification, but this standard cannot distinguish between different populations of Chinese soft-shelled turtle.

The identification methods for conventional detection of different populations of soft-shelled turtles include morphological marker identification, isozyme marker identification and mtDNA mitochondrial sequence identification, etc. However, the external morphological marker identification method is greatly affected by the breeding environment of Chinese soft-shelled turtle, and is prone to misjudgment; Markers have tissue specificity and individual differences, and are affected by individual development stages, so they cannot meet high accuracy requirements; the RAPD method has poor repeatability and high template requirements, so the reliability of the identification results is also limited; mtDNA mitochondria Although the sequence identification method is accurate, it needs to be completed with the help of a large-scale sequencer. Most laboratories are often unable to purchase the instrument due to the high price, so they can only send the amplified sample out for sequencing and then conduct germplasm identification. Timeliness cannot ensure

According to domestic and foreign literature search, there is no relevant report on stable, reliable and simple germplasm identification of four different populations of soft-shelled soft-shelled turtle

Images