Method for detecting base mutation polymorphism of goat gonadotropin releasing hormone and growth differentiation factor 9

Abstract

The invention discloses a method for detecting the polymorphism of three base mutations of the gene exon 4 of goat gonadotropin releasing hormone and the gene exon 2 of the growth differentiation factor 9. The method comprises the following steps: using the DNA sequence of goat genome as template, using 2 to amplify the primer P1 and P2 under PCR condition in the presence of Taq DNA polymerase, buffer environment, Mg2+ and dNTPs, using agarose gel electrophoresis to judge the size of the target fragments; using polyacrylamide gel electrophoresis to detect the PCR amplified product of the primer P1 and P2 to find that the two amplification sites have three base mutations, then performing genotyping and gene frequency analysis to the SNPs of the two genes, and performing association analysis with the fecundity traits of Boer goat and Xinong Saanen dairy goat. The result shows that the SNPs sites detected by the primer P1 and P2, of the GnRH gene exon 4 (containing partial intron 3 and 3' non-translational region) and the GDF9 gene exon 2, thus the method can be used for molecular marking of fecundity trait selection of goat.

Description

technical field

[0001] The invention belongs to the field of molecular genetics, and in particular relates to a three-way test for detecting goat gonadotropin-releasing hormone gene exon 4 (including part of intron 3 and 3' untranslated region) and growth differentiation factor 9 gene exon 2 A base mutation polymorphism method, which uses polyacrylamide gel electrophoresis (PAGE), has the characteristics of rapidity, accuracy and simplicity. Background technique

[0002] Single Nucleotide Polymorphisms (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 ...

Examples

Embodiment Construction

[0015] , GnRH gene exon 4 (including part of intron 3 and 3'untranslated region) and GDF9 gene exon 2 PCR amplification and polymorphism detection 1. Collection and processing of goat blood samples Take goat blood samples Add 0.2 μL of ACD (citric acid 2.4g; trisodium citrate 6.6g; glucose 7.35g; dilute to 50mL, autoclave) to anticoagulate, slowly invert 3 times, put in ice box, and store at -80°C spare.

[0016] In this example, a total of 720 ewe blood samples from 2 goat populations were used, specifically: 406 blood samples of Xinong Saanen sheep, collected from Qianyang Saaneng sheep breeding farm in Shaanxi Province; 314 blood samples of Boer goats, collected from Shaanxi Province Boer Goat Breeding Farm in Linyou County and Luonan Branch of Yangling Jinkun Company in Shaanxi Province.

[0017] 2. Extraction and purification of genomic DNA from blood sample 1) Thaw the frozen blood sample at room temperature, transfer 500 μL to a 1.5 mL Eppendorf tube, add an equal volu...

technical field

[0001] The invention belongs to the field of molecular genetics, and in particular relates to a three-way test for detecting goat gonadotropin-releasing hormone gene exon 4 (including part of intron 3 and 3' untranslated region) and growth differentiation factor 9 gene exon 2 A base mutation polymorphism method, which uses polyacrylamide gel electrophoresis (PAGE), has the characteristics of rapidity, accuracy and simplicity. Background technique

[0002] Single Nucleotide Polymorphisms (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 ...

Examples

Embodiment Construction

[0015] , GnRH gene exon 4 (including part of intron 3 and 3'untranslated region) and GDF9 gene exon 2 PCR amplification and polymorphism detection 1. Collection and processing of goat blood samples Take goat blood samples Add 0.2 μL of ACD (citric acid 2.4g; trisodium citrate 6.6g; glucose 7.35g; dilute to 50mL, autoclave) to anticoagulate, slowly invert 3 times, put in ice box, and store at -80°C spare.

[0016] In this example, a total of 720 ewe blood samples from 2 goat populations were used, specifically: 406 blood samples of Xinong Saanen sheep, collected from Qianyang Saaneng sheep breeding farm in Shaanxi Province; 314 blood samples of Boer goats, collected from Shaanxi Province Boer Goat Breeding Farm in Linyou County and Luonan Branch of Yangling Jinkun Company in Shaanxi Province.

[0017] 2. Extraction and purification of genomic DNA from blood sample 1) Thaw the frozen blood sample at room temperature, transfer 500 μL to a 1.5 mL Eppendorf tube, add an equal volu...

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