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Primers, kit and detection method for identifying carcasses of silky chicken and black-bone chicken

A technology for black phoenix chickens and silky chickens, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem that there is no effective method for identifying silk-feathered silky chickens and black phoenix chicken carcasses. and other issues, to achieve the effect of good social value, broad market application prospects and considerable economic benefits

Active Publication Date: 2021-06-08
NANCHANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no effective method to identify the carcasses of silky and black-bone chickens

Method used

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  • Primers, kit and detection method for identifying carcasses of silky chicken and black-bone chicken
  • Primers, kit and detection method for identifying carcasses of silky chicken and black-bone chicken
  • Primers, kit and detection method for identifying carcasses of silky chicken and black-bone chicken

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Identification of genome differences between silky silky chicken and black phoenix chicken

[0029] Step 1: Template Preparation

[0030] (1) Tissue collection: The skin samples of 24 silky chickens and 24 black-bone chickens were collected.

[0031] (2) DNA template preparation: about 50 mg of skin samples were collected, put into a centrifuge tube with 500 μL of liquid nitrogen, and the collected samples were ground with a tissue grinder. After the liquid nitrogen was evaporated to dryness, 200 microliters of TE was added, centrifuged at 1200 rpm / min, and the supernatant was extracted for later use.

[0032] Step 2: Type the target locus

[0033] (1) PCR amplification

[0034] PCR amplification primers are shown in Table 1:

[0035] Table 1 PCR amplification primers

[0036]

[0037] Note: "S1-CXF" is the sequencing primer corresponding to the PCR amplified fragment. Using additional primers for sequencing can save the step of PCR product purificati...

Embodiment 2

[0056] Reliability of Embodiment 2 Single-blind Detection and Judgment Method

[0057] In this example, a single-blind method is adopted to determine the source of the species of the samples of known species according to the molecular detection results.

[0058] Step 1: Template Preparation

[0059] (1) Tissue collection: The skin tissues of 12 silky chickens and 12 black-bone chickens were collected.

[0060] (2) DNA template preparation: about 50 mg of skin samples were collected, put into a centrifuge tube with 500 μL of liquid nitrogen, and the collected samples were ground with a tissue grinder. After the liquid nitrogen was evaporated to dryness, 200 microliters of TE was added, centrifuged at 1200 rpm / min, and the supernatant was extracted for later use.

[0061] Step 2: Type the target locus

[0062] (1) PCR amplification

[0063] PCR amplification primers are shown in Table 7:

[0064] Table 7 PCR amplification primers

[0065]

[0066] Note: "S1-CXF" is the se...

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Abstract

The invention discloses primers, kit and detection method for identifying carcasses of a silky chicken and a black-bone chicken. The primers include a forward primer with a nucleotide sequence as shown in SEQ ID No.1, a reverse primer with a nucleotide sequence as shown in SEQ ID No.2 and a sequencing primer with a nucleotide sequence as shown in SEQ ID No.3. On the basis of polymorphism of a PCR product sequence base site of a chicken breed to be detected, allelic genes of the site are judged through a gene sequencing method; when a base at 161 bp is C, a base at 223 bp is A, a base at 347 bp is T, a base at 376 bp is A, a base at 586 bp is T and / or a base at 593 bp is A, it is judged that the chicken breed is a black-bone chicken breed; and when the bases do not appear at the site, it is judged that the chicken breed is the silky chicken. According to the method, the carcasses of the silky chicken and the black-bone chicken can be accurately judged by 100%.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer, a kit and a detection method for identifying carcasses of silky chickens and black phoenix chickens. Background technique [0002] Silky chicken with silk feathers is a special product of Taihe County, Jiangxi Province. It is native to the northern foot of Wushan Mountain in Taihe County. It is famous for its ten characteristics of "claw, black skin, black meat, and black bone" as well as its high nutritional value and medicinal value. Silkie chicken is a famous food and medicine chicken. The whole body can be used as medicine. The bones, meat and viscera have medicinal value, and can be formulated into various patent medicines and prescriptions. [0003] However, the original silky-bone chicken grows slowly and has low egg production, so commercial breeding companies use silky-feather silky and fast-growing breeds as breeding materials to breed black phoenix...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 许继国崔芳芳周敏杨艳北贡继尚李袁飞朱学农谭玉文熊信威许桥马荆鄂盛文涛饶友生王樟凤
Owner NANCHANG NORMAL UNIV
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