Method for detecting mutation polymorphism of 5 basic groups in gene coding region of goat growth hormone

Abstract

The invention discloses a polyacrylamide gel electrophoresis method for detecting the mutation polymorphism of 5 basic groups in gene coding region of a goat growth hormone (GH). The method comprises the following steps of: performing amplification under a PCR condition by using two pairs of reaction (PCR) primers P1 and P2 in the presence of a goat genome DNA sequence as a template, TaqDNA polymerase, Buffer (a buffer environment), Mg2<+> and dNTPs; determining the sizes of amplification objective fragments according to an agarose gel electrophoresis; screening the mutation of the 5 basic groups in the gene coding region of the goat GH by using a DNA sequencing technique, and then performing genotyping and gene frequency analysis on the SNPs of a GH gene; and performing association analyses on the growth traits of a Boer goat and three-moon's age filial generations (F1 and F2) of the Boer goat and a Guanzhong dairy goat, wherein the results of the show that the detected SNPs sites of the P1 and the P2 of a GH gene can be used as molecular markers of the early selection (three-moon's age) of the growth traits of a goat.

Description

technical field [0001] The invention belongs to the field of molecular genetics, in particular to a method for detecting goat growth hormone ( GH ) method for 5 base mutation polymorphisms in the gene coding region. Background technique [0002] SNP (Single Nucleotide Polymorphisms, SNP) refers to the polymorphism caused by the replacement of a single nucleotide (A / T / C / G) in the genomic DNA sequence. SNPs can be polymorphisms of two or more alleles. Therefore, commonly referred to as SNPs include base insertions, deletions, insertions / deletions, and changes in the copy number of repeated sequences. A SNP represents a nucleotide change at a certain position in the genome, mainly consisting of single base conversions (one pyrimidine for another or one purine for another) and transversions (purine exchanged with pyrimidine). SNPs with transversion variants account for about 2 / 3, and several other SNPs are at similar levels, because CpG (representing nucleotide pairs, where G...

AI Technical Summary

Problems solved by technology

Among these SNP detection techniques, DNA sequence determination is the most accurate SNP detection method, but its detection cost is extremely expensive, and large-scale instruments such as DNA sequencers are required. At the same time, very skilled technicians and Experience, therefore, DNA sequencing is not an ideal SNP detection method for practical production

Of course, the combination of PCR-SSCP and DNA sequencing to detect SNP can reduce the detection cost appropriately, but the experimental process of PCR-SSCP is relatively long, the operation is cumbersome, and there are false negative problems in the experimental process, so it is not an ideal SNP Detection method; as a new type of SNP detection method, the AS-PCR method has very broad prospects in the future application field, but this method requires the design of special primers and can only target specific gene loci. At the same time, There is also a probability of false detection in the detection process, so it does not have the characteristics of general application at present; while the detection of SNP sites by primer extension method and oligonucleotide ligation reaction technology requires plate reader, gene chip, microsphere array technology and Detection platforms such as mass spectrometers are not very practical for general molecular laboratories

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