Kit and method for detecting A746G mutation of sheep BMPR-IB gene by using Taqman-MGB probe

A probe detection and detection method technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of difficult optimization of reaction conditions and troublesome operation, and achieve good probe quenching effect, Low cost, effect of reducing synthesis cost

Active Publication Date: 2017-11-10
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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Problems solved by technology

ARMS-PCR method for detecting SNP sites is cumbersome to operate and difficult to optimize reaction conditions

Method used

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  • Kit and method for detecting A746G mutation of sheep BMPR-IB gene by using Taqman-MGB probe
  • Kit and method for detecting A746G mutation of sheep BMPR-IB gene by using Taqman-MGB probe
  • Kit and method for detecting A746G mutation of sheep BMPR-IB gene by using Taqman-MGB probe

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Embodiment 1

[0042] Reference attached Figure 1-Figure 3 , the technical scheme of the present invention is described in detail below in conjunction with accompanying drawing embodiment, the implementation steps of present embodiment are as follows:

[0043] 1. Experimental materials and sample collection:

[0044] Select 498 Chinese Merino (Military Reclamation Type) multiple-birth lines for meat, 464 Chinese Merino (Military Type) multiple-birth lines for wool, 663 multiple-birth Suffolk sheep, 150 Suffolk sheep, and 320 Kazakh sheep and 150 Hu sheep as the experimental subjects, a total of 2245.

[0045] Draw about 1 mL of blood from the jugular vein of sheep with a disposable syringe, inject it into a 1.5 mL centrifuge tube that has been autoclaved and filled with about 150 µL of 2% sterile EDTA (Ethylene diamine tetraacetic acid, EDTA) anticoagulant, and shake gently Mix well, record the number of sheep, and store at -20°C for later use.

[0046] 2. Main drugs and reagents:

[00...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a kit and method for detecting the A746G mutation of a sheep BMPR-IB gene by using a Taqman-MGB probe. The kit mainly comprises three tubes of positive control samples, a DNA extraction solution I, a DNA extraction solution II, a 2 * Light Cycler 480 probes master reaction buffer solution, BMF and BMR primers, BPA and BPG probes and ddH2O. The Method includes the steps of rapidly extracting genomic DNA, conducting real-time fluorescent quantitative PCR amplification on the genomic DNA by using the primers, and performing gene interpretation. The kit and method for detecting the A746G mutation of the sheep BMPR-IB gene by using the Taqman-MGB probe has the advantages of being simple to operate, fast in detection and high in accuracy, and can judge types without subsequent analysis of PCR products; besides, the whole reaction process is performed in a closed tube, so that cross-contamination is reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Taqman-MGB probe detection kit and method for the A746G mutation of sheep BMPR-IB gene. Background technique [0002] High fecundity is one of the important goals pursued by sheep production in the world. However, because most of the sheep produce single lambs and a few produce double lambs, the reproductive performance of sheep is greatly affected. key. The fecundity trait is one of the important reproductive traits of sheep, and it is also the basis of high-yield and high-yield sheep. The level of fecundity directly affects the production cost, and then affects the economic benefits. Or breed multiple litter sheep breeds. Studies have shown that most sheep breeds have a low number of lambs, and the heritability of multiple lambs is low, only 0.12, which limits the improvement of sheep production performance in my country to a certain extent. It affects the effect a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/172C12Q2561/101C12Q2545/113C12Q2563/107
Inventor 杨华杨永林李良远卢守亮杨涵羽璐
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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