Kit for detecting reproductive capacity of sheep and using method thereof

A kit and capability technology, which is applied to the kit for detecting the reproductive ability of sheep and its application field, can solve the problems of long detection cycle, inappropriateness, and high detection cost, and achieve the reduction of DNA extraction cost, simple and easy operation of SNP typing, The effect of improving the speed of DNA extraction

Inactive Publication Date: 2011-08-24
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing technology, only one gene SNP can be detected at a time, so the cost of each gene SNP detection is very high, and it is estimated that the general cost is about 13 US dollars/sheep
At the same time, the method of the prior art is complicated to

Method used

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  • Kit for detecting reproductive capacity of sheep and using method thereof
  • Kit for detecting reproductive capacity of sheep and using method thereof
  • Kit for detecting reproductive capacity of sheep and using method thereof

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specific Embodiment approach

[0019] FecB and FecX G Preparation of Gene Cloning Standards and Genotyping Detection

[0020] (1). The primer gene sequences and probe sequences used in the present invention listed in Table 1 and Table 2 below were all synthesized by Shanghai Sangon Bioengineering Technology Development Co., Ltd.

[0021] Table 1. FecB and FecX G Gene Cloning Primers

[0022]

[0023] (2) FecB and FecX G Gene construction and sequencing

[0024] The synthesized FecB(AA), FecB(GG), FecX G (CC) and FecX G (TT) Genotype double-stranded nucleic acid sequences were amplified by PCR with F1 / R1 and F2 / R2 primers respectively, followed by 1% agarose gel electrophoresis, and 187bp and 87bp fragments were recovered from the gel, and the recovered PCR products were used T4 ligase Ligated with pMD19-T cloning vector to construct pMD19-T FecB(AA), pMD19-T FecB(GG), pMD19-T FecX G (CC) and pMD19-T FecX G (TT) vector, transformed Escherichia coli (Escherichia.coli) competent cell JM109. Positiv...

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Abstract

The invention discloses a kit for detecting the reproductive capacity of sheep and a using method thereof. The kit is internally provided with primer nucleotide sequences shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4 in a sequence table, probe nucleotide sequences shown in SEQ ID NO:5 and SEQ ID NO:6 as well as respectively placed FTA (Fault Tree Analysis) card and relevant reagents. The using method of the kit provided by the invention comprises the following steps: adding the primer nucleotide sequences, probe nucleotide sequences and relevant reagents in the kit into the DNA (Deoxyribose Nucleic Acid) of the sheep to be detected; then carrying out PCR (Polymerase Chain Reaction) amplification; carrying out fusion curve analysis on an amplification product in a high-resolution fusion curve analyzer; and forecasting the lambing situation of the sheep to be detected according to G, A, T and C peaks in the fusion curve.

Description

technical field [0001] The invention relates to a detection kit and its application method, in particular to a detection kit for detecting sheep reproductive ability and its application method. Background technique [0002] The reproductive traits of sheep are important economic traits of sheep, which are quantitative traits and low heritability traits. It includes three aspects: conception rate, multiple fecundity and lamb survival rate. Among them, multiple fecundity is one of the most important traits that determine the benefits of sheep farming, and it is also the basis for high-yield and high-yield sheep industries. At present, the world's major sheep producing countries have attached great importance to the study of sheep's fecundity performance. The economic benefits of raising sheep will be greatly improved through the detection of sheep reproductive traits. [0003] At present, the fundamental means to improve sheep fecundity is genetic selection, but because the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 岳耀敬杨博辉牛春娥冯瑞林郎侠刘建斌孙晓萍郭建郭婷婷
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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