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Kit for detecting specific microsatellite locus of sheep Y chromosome and application of kit

A technology of microsatellite loci and detection kit, applied in the field of biology, can solve the problem of ignoring genetic effects and other problems

Inactive Publication Date: 2014-12-17
王玉涛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Autosomal microsatellite and mtDNA markers study the origin of domesticated animals, ignoring paternal genetic effects, with certain limitations

Method used

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  • Kit for detecting specific microsatellite locus of sheep Y chromosome and application of kit
  • Kit for detecting specific microsatellite locus of sheep Y chromosome and application of kit
  • Kit for detecting specific microsatellite locus of sheep Y chromosome and application of kit

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Experimental program
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Effect test

Embodiment Construction

[0017] The technical solutions of the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.

[0018] Blood genomic DNA extraction (using traditional phenol-chloroform extraction method)

[0019] (1) After the frozen blood is thawed at room temperature, transfer 500 μl to a centrifuge tube, add an equal volume of PBS buffer, mix, shake well, centrifuge at 12,000 rpm for 10 min, and discard the supernatant.

[0020] (2) Take another 500 μl of blood and transfer it to a centrifuge tube, add an equal volume of PBS buffer, mix, shake well, centrifuge at 12,000 rpm for 10 min, and discard the supernatant.

[0021] (3) 500 μl of DNA extraction buffer (20% SDS) was added, and the precipitate was resuspended.

[0022] (4) Add proteinase K to a final concentration of 100 μg / ml, mix well, seal with parafilm, and water bath at 55°C overnight.

[0023] (5) Cool the solution to room temperature, add an equal volum...

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Abstract

The invention discloses a kit for detecting a specific microsatellite locus of a sheep Y chromosome and an application of the kit. The kit comprises 0.6 microliter of to-be-detected sample template DNA, 0.2 microliter of specific microsatellite forward primer, 0.2 microliter of reverse primer, 1 microliter of nucleic acid amplification system 6*Buffer, 0.8 microliter of dNTP, 0.2 microliter of TaqDNA polymerase, and finally adding ddH2O to obtain 10 microliters of mixture. A microsatellite locus is screened out from a specific region of the sheep Y chromosome, and a product can be amplified when the locus is applied to male sheep, but no product is amplified when the locus is applied to a female sheet body. The male sheep group has certain polymorphism by further research, which shows that the locus can be subjected to early embryo sex identification of the sheep, and can also be applied to paternal origin evolution research of the sheep.

Description

technical field [0001] The invention belongs to the technical field of biology, and relates to a sheep Y chromosome-specific microsatellite site detection kit and application thereof. Background technique [0002] Autosomal microsatellites and mtDNA markers to study the origin of domestic animals, ignoring the genetic effects of the paternal parent, have certain limitations. In recent years, many researchers such as Underhill P A have realized that genetic markers on the Y chromosome are ideal tools for studying the origin, evolution, and migration of species. As paternally inherited chromosomes, most of them are non-recombination regions, so the haplotype remains intact and is not easily affected by recombination and reversion. The mutation rate is low, and it is more stably inherited than microsatellite markers. It is a faithful recorder of evolutionary events. Furthermore, since chromosomes are haploid, their effective population size is much smaller than that of autosom...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6879C12Q2600/156
Inventor 王玉涛韩建林罗玉柱郭丽君
Owner 王玉涛
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