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Method of authenticating chicken contour feathers and silken feathers by applying fluorescence probe PCR (polymerase chain reaction)

A fluorescent probe and identification method technology, which is applied in the field of chicken molecular marker preparation, can solve the problems of high production cost and long breeding time, and achieve the effects of short detection time, strong specificity and high detection sensitivity

Inactive Publication Date: 2015-12-23
GUANGXI ZHUANG AUTONOMOUS REGION INST OF ANIMAL HUSBANDRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a method for identifying chicken flake feathers and silk feathers using fluorescent probe PCR in order to overcome the defects of long breeding time and high production costs in the conventional breeding of poultry feather traits

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  • Method of authenticating chicken contour feathers and silken feathers by applying fluorescence probe PCR (polymerase chain reaction)
  • Method of authenticating chicken contour feathers and silken feathers by applying fluorescence probe PCR (polymerase chain reaction)
  • Method of authenticating chicken contour feathers and silken feathers by applying fluorescence probe PCR (polymerase chain reaction)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 : Design and synthesis of primers and Taqman probes

[0038] A pair of specific primers and two Taqman probes (Table 1) were designed by using the PrimerExpress 5.0 software according to the DNA sequence of chicken chromosome 3 in Ganbank and chicken feather traits.

[0039] Table 1 Primer and Taqman probe sequence (5'-3')

[0040]

Embodiment 2

[0041] Example 2: Fluorescent probe PCR detection

[0042] 2.1 Extraction of nucleic acid

[0043] The DNA of GG genotype, CC genotype and GC genotype was extracted according to the instructions of the blood genome column extraction kit.

[0044] 2.2 Preparation of standard genotypes

[0045] PCR amplification was performed with self-designed primers, the upstream primer was 5'-GACTCTCAACGCGGGAAC-3', and the downstream primer was 5'-CTGGGGGCAGCCATCTT-3'.

[0046] A 25 μL reaction system was used: 2×EsTaqMasterMix 15 μL, forward and reverse primers 0.5 μL each, DNA template 2 μL, and water to 25 μL.

[0047] The PCR conditions are: first step: 95°C for 5min, second step: 95°C for 30s, third step: 62°C for 30s, fourth step: 72°C for 30s, fifth step: 72°C for 5min; Four-step cycle 30 times.

[0048] The PCR product was detected by 1% agarose gel electrophoresis, and the target gene was recovered and ligated with pGM-Tvector to obtain a recombinant plasmid.

[0049] The lig...

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Abstract

The invention discloses a method of authenticating chicken contour feathers and silken feathers by applying a fluorescence probe PCR (polymerase chain reaction). The authenticating method is completed by using a fluorescence probe PCR primer group, a reagent I of the fluorescence probe PCR and a reagent II of the fluorescence probe PCR. The method disclosed by the invention has the advantages of short detection time, high detection sensitivity and strong specificity.

Description

technical field [0001] The invention belongs to the technical field of chicken molecular marker preparation. Specifically relates to a method for identifying chicken flake feathers and silk feathers by using fluorescent probe PCR. Background technique [0002] At present, the breeding method of new breeds of poultry is mainly selective breeding. Many new breeds have been bred by means of selective breeding. However, the selection of quantitative traits in conventional breeding methods can only be manifested when the individual grows and develops to a certain stage, and the breeding cycle is long. The problem of high production cost has seriously restricted the rapid development of poultry breeding industry. [0003] The feather traits of poultry are mainly silk feathers and sheet feathers, which are determined by a single nucleotide polymorphism (SNP) site on chicken chromosome 3, which is located on ss666793747 / rs316090093. At this locus, the feathers of the GG and GC gen...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/172C12Q2563/107C12Q2561/101
Inventor 莫国东黄英飞廖玉英吴强覃仕善韦凤英
Owner GUANGXI ZHUANG AUTONOMOUS REGION INST OF ANIMAL HUSBANDRY