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Molecular biological variety identification method for Huangjiaji chicken

A molecular biology and identification method technology, applied in the field of biological species identification, can solve problems such as difficult, unscientific, accurate, and distinguishable, and achieve the effects of low cost, short identification time, and high accuracy

Inactive Publication Date: 2012-03-21
MODERN NEW AGRI GROUP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the continuous maturity of breeding technology, the appearance characteristics of the parents and the hybrid offspring are quite similar, and it is difficult to distinguish them from the appearance characteristics.
Therefore, it is not scientific and accurate enough to use morphological markers to identify poultry breeds (strains).
If the production performance measurement is carried out, it will not only consume a lot of manpower, time and high performance measurement costs, but also the results of the performance measurement are also affected by factors in various aspects of feeding, so the production performance measurement is also difficult to accurately reflect the true nature of each variety (strain). Production performance and germplasm characteristics

Method used

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  • Molecular biological variety identification method for Huangjiaji chicken
  • Molecular biological variety identification method for Huangjiaji chicken
  • Molecular biological variety identification method for Huangjiaji chicken

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The tested chicken is Huangjiaji chicken:

[0031] (1) Extract the total DNA including mitochondrial DNA from Huangjiaji chicken (1, breed code named HJJ1) and tested chicken (2 Huangjiaji chickens, breed code named SHJJ1 and SHJJ2) respectively by phenol-chloroform method;

[0032] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:

[0033] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';

[0034] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.

[0035] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';

[0036] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.

[0037] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';

[0038] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.

[0039] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplif...

Embodiment 2

[0055] The tested chicken is native chicken:

[0056] (1) Extract the total DNA of Huangjiaji chicken (1, breed code-named HJJ1) and tested chickens (3 native chickens, breed code-named TJ1, TJ2 and TJ3) including mitochondrial DNA by the phenol-chloroform method;

[0057] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:

[0058] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';

[0059] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.

[0060] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';

[0061] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.

[0062] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';

[0063] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.

[0064] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplified product is cl...

Embodiment 3

[0077] The tested chicken is black-bone chicken:

[0078] (1) Extract the total DNA of Huangjiaji chicken (1, breed code-named HJJ1) and tested chickens (3 black-bone chickens, breed code-named WJ1, WJ2 and WJ3) including mitochondrial DNA by the phenol-chloroform method;

[0079] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:

[0080] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';

[0081] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.

[0082] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';

[0083] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.

[0084] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';

[0085] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.

[0086] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplified product...

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Abstract

The invention discloses a molecular biological variety identification method for a Huangjiaji chicken, which comprises the following steps: accurately acquiring COI gene sequences of mitochondrion DNAs of a Huangjiaji chicken and a tested chicken based on gene cloning and DNA sequencing technologies, calculating variable sites, haplotypes and average nucleotide differences, nucleotide degrees of ramification and net genetic distances of the Huangjiaji chicken and the tested chicken by means of DnaSP4.10.7 software, constructing colony clustering charts and haplotype phylogenetic trees of the Huangjiaji chicken and the tested chicken, and determining whether the Huangjiaji chicken and the tested chicken are of the same variety according to the results of the constructed colony clustering charts and haplotype phylogenetic trees. By identifying whether the Huangjiaji chicken is true or false based on the precise software calculation and analysis results of the differences between varieties in genetic information, the invention has the characteristics of high accuracy, short identification time, low cost and the like.

Description

technical field [0001] The invention belongs to the technical field of biological variety identification, and in particular relates to a molecular biological identification method of Huangjiaji chicken variety. Background technique [0002] Huangjiaji chicken is a high-quality new breed of broiler chicken newly developed by Modern New Agriculture Co., Ltd. It is widely favored by consumers because of its delicious meat, mellow taste and juicy taste. However, some unscrupulous enterprises and farmers in the market illegally produce and sell counterfeit and counterfeit Huangjiaji chickens, which has seriously damaged the reputation of the Huangjiaji chicken brand and affected the development of Modern New Agriculture Co., Ltd. At present, the identification of poultry breeds is mainly based on morphological markers to review breeding files and performance measurements. However, with the continuous maturity of breeding technology, the appearance characteristics of the parents ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 宋金典
Owner MODERN NEW AGRI GROUP
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