Breeding method of multi-gene pyramiding of fine breed milk goats

A milk goat and polygene technology, which is applied in the breeding field of multi-gene aggregation of fine-bred dairy goats, can solve the problems of expensive detection costs, cumbersome operations, and poor implementation of molecular laboratories, so as to speed up the progress of breeding and improve the efficiency of selection Effect

Inactive Publication Date: 2010-09-29
NORTHWEST A & F UNIV
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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, so DNA sequence determination is not an ideal SNP detection method for actual production; of course, the combination of PCR-SSCP and DNA sequencing can be used to detect SNPs can appropriately reduce the detection cost, but the experimental process of PCR-SSCP is relatively long , the operation is cumbersome, and there are false negative problems in...

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  • Breeding method of multi-gene pyramiding of fine breed milk goats
  • Breeding method of multi-gene pyramiding of fine breed milk goats
  • Breeding method of multi-gene pyramiding of fine breed milk goats

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[0022] A. PCR amplification of PRLR intron 2 and exon 10 using primers P1 and P2, and PCR amplification and polymorphism of LHβ gene 5'UTR and exon 1 using primers P3 and P4 1. Goat Blood sample collection and processing Take 5 mL of goat blood sample, add 0.2 μL of ACD (2.4 g of citric acid; 6.6 g of trisodium citrate; 7.35 g of glucose; dilute to 50 mL, autoclave) for anticoagulation, slowly invert 3 times and put Store in an ice box at -80°C for later use.

[0023] In this example, 265 blood samples of Xinong Sanen dairy goats that had continuously produced 2 or more lambs were collected from Qianyang Sanen Sheep Breeding Farm in Shaanxi Province and Shaanxi Green New Century Biological Co., Ltd.

[0024] 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 volume of PBS buffer, mix well, centrifuge at 12000 r / min for 10 min (4 ° C), discard the supernata...

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Abstract

The invention discloses a breeding method of multi-gene pyramiding of fine breed milk goats, which comprises the following steps: using genome DNA of milk goats continuously bearing two or more than two goats as a template; using four pairs of primers for respectively amplifying intrones 2 and exons 10 of prolactin receptor genes, and the 5' non-translational region (5' UTR) and exons 1 of luteotropin beta subunit genes; using agarose gel electrophoresis for carrying out size determination on each amplifying product; adopting the DNA sequencing technology for sieving and checking the site-directed mutation of the amplifying products of the four pairs of the primers; then, using polyacrylamide gel electrophoresis for carrying out gene typing and gene frequency analysis on the SNPs of the four sites of the prolactin receptor genes and the luteotropin beta subunit genes; analyzing the relationship between the goat bearing number and the polymorphism of the high-yield milk goat individuals (generation F1) and the relationship between different gene type combinations and the goat bearing number; tracing to the parent generation (generation F0); tracking filial generation (generation F2); detecting the relationship between the goat bearing number and the gene type combinations of the female goat individuals; and analyzing the contribution of different gene types in the prolific trait formation.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and in particular relates to a breeding method for multi-gene polymerization of fine-bred dairy goats. The method uses PCR-SSCP and DNA sequencing techniques to detect prolactin receptor (PRLR) genes and luteinizing hormone beta Single-strand DNA base mutation polymorphism of subunit (LHβ) gene, determine the appropriate molecular marker by analyzing the relationship between polymorphism and litter size of high-yield dairy goat individuals (F1 generation), and analyze high-yield dairy goat individuals (F1) The relationship between the different genotype combinations of ewes and the number of litters, trace the parents (F0 generation), track the offspring (F2 generation), detect the relationship between the genotype combinations of individual ewes and the number of litters, and study the relationship between different genotypes in Then select the genotype combinations of high-yielding individuals, ...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 曹斌云安小鹏李广侯金星王建刚宋宇轩杨明明朱广琴王韵斐崔易虹陈秋菊
Owner NORTHWEST A & F UNIV
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