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PCR kit used for detecting polymorphism of sheep BMPR-IB genes

A gene polymorphism and kit technology, applied in the fields of molecular biology and animal breeding, can solve the problems of time-consuming, labor-intensive, high consumption, and low degree of automation.

Inactive Publication Date: 2012-10-17
HEBEI AGRICULTURAL UNIV.
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the detection of sheep multifetal genes, the common method is PCR-RFLP. After PCR, enzyme digestion and electrophoresis are time-consuming and labor-intensive.
[0004] This work involves a lot of repetitive labor, high consumption, and low degree of automation. This method is not suitable for large-scale, low consumption, and automation requirements. Therefore, it is potential to establish an automated, simple, and fast sheep BMPR-IB genotyping method. must do

Method used

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  • PCR kit used for detecting polymorphism of sheep BMPR-IB genes
  • PCR kit used for detecting polymorphism of sheep BMPR-IB genes
  • PCR kit used for detecting polymorphism of sheep BMPR-IB genes

Examples

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Effect test

Embodiment

[0048] The test kit in this embodiment is made up of separately packaged primer primer F, primer R, probe Anchor, probe Sensor and PCR reaction solution, wherein

[0049] The sequences of primers F and R are:

[0050] F5-AGAGGACAATAGCAAAGCA-3:

[0051] R5-TGAACATCGCTAATACAGACTT-3:

[0052] The probe sequence is:

[0053] Anchor probe: AGGCCAGCTGGTTCCGAGAGA-BlackHQ;

[0054] Sensor probe: FAM-AGAAATATATCAGACGGTGTTG-PO4

[0055] On the probe Anchor and sensor, BlackHQ is a quencher, and FAM is a green fluorescent marker.

[0056] The kit in this example was prepared based on a 25ul reaction system, and the kit contained 12.5ul probe mixPCR reaction solution, 0.2uM primer F, 1uM primer R, 0.2uMAnchor probe, and 0.1uM Sensor probe. When testing, take 1ul DNA (20-50ng) sample and add it to the above reaction system.

[0057] In order to further illustrate the use method and use effect of the kit made in this example, the specific process and use effect of using the above kit ...

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Abstract

The invention provides a PCR kit used for detecting polymorphism of sheep BMPR-IB genes. The kit comprises separately packaged primer F, primer R, Anchor probe, Sensor probe and PCR reaction solution, wherein the sequences of the primer F and the primer R are F5-AGAGGACAATAGCAAAGCA-3 and R5-TGAACATCGCTAATACAGACTT-3 respectively and the sequences of the Anchor probe and the Sensor probe are AGGCCAGCTGGTTCCGAGAGA-BlackHQ and FAM-AGAAATATATCAGACGGTGTTG-PO4 respectively.

Description

technical field [0001] The present invention relates to the fields of molecular biology and animal breeding. Specifically, it relates to single nucleotide polymorphism (Single nucleotide polymorphism, abbreviated as SNP) of BMPR-IB gene and its correlation with lamb size of sheep, and relates to a kit for detecting these SNPs. Background technique [0002] The study found that the multifetal performance of Booroola Merino sheep is caused by the mutation of the FecB gene located on the autosome. The mutation of the FecB gene has an additive effect on the ovulation rate of sheep, and a partial dominant effect on the litter size. The sheep FecB gene is actually the bone morphoprotein type IB receptor (BMPR-IB) gene. Due to the mutation of the A746G base of the gene, the glutamic acid at position 249 is mutated to arginine (Q→R), resulting in the number of ovulations and The litter size increased, and it proved that the BMPR-1B gene was the main gene affecting the litter size o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 张英杰刘月琴刘彦琴王红娜孙洪新董李学李雪梅杨佳栋
Owner HEBEI AGRICULTURAL UNIV.
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