A kind of molecular identification primer and identification method of fast and slow feather phenotype of chicken

A technology of molecular identification and speed, which is applied in the field of molecular genetics, can solve the problems of long time for establishment of male and female matching lines, limited identification time, and unclear identification standards, etc., and achieves chicken blood processing and preservation conditions are simple, easy to operate, The effect of avoiding human error

Inactive Publication Date: 2018-06-15
HEBEI AGRICULTURAL UNIV. +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The second object of the present invention is to provide a molecular identification method for the fast and slow feather phenotype of chickens, which can solve the problems that the identification standards in the identification of fast and slow feathers are not clear enough, the establishment of male and female matching lines takes a long time, and the identification time is limited.

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  • A kind of molecular identification primer and identification method of fast and slow feather phenotype of chicken
  • A kind of molecular identification primer and identification method of fast and slow feather phenotype of chicken
  • A kind of molecular identification primer and identification method of fast and slow feather phenotype of chicken

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Embodiment 1

[0041] A molecular identification method for chicken fast and slow feather phenotype, comprising the following steps:

[0042] 1), template DNA extraction: usually extracted from chicken blood, of course, can also be extracted from other tissues;

[0043] 2), use the primers shown in SEQ ID NO:1 and SEQ ID NO:2, and carry out PCR amplification with the DNA extracted in 1) as a template;

[0044] 3), carrying out agarose gel electrophoresis to the product obtained by PCR amplification in 2);

[0045] 4). Analyze the electrophoresis results in 3).

Embodiment 2

[0047] 1. Primer preparation

[0048] The primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., in which:

[0049] Forward primer 1305s: 5'CCTCCTTGCCAAACCTTA3';

[0050] Reverse primer 1305a: 5'ATCAGCCAGATCCGTCAG3'.

[0051] 2. The kit extracts chicken blood clot DNA steps:

[0052] The ClotBlood DNA Kit from Kangwei Century Biotechnology Co., Ltd. was selected for extraction, product number CW0545.

[0053] (1) Weigh 0.1-0.2 g of coagulated blood clot, place it in a centrifuge tube, add 550 μl Buffer GTL, and homogenize with an electric homogenizer. In order to fully crush the blood clot, small magnetic beads can be added for mechanical crushing.

[0054] (2) Add 20 μl of proteinase K and invert the sample upside down to fully mix the sample. Incubate overnight at 56°C in a shaker until all particles are completely dissolved.

[0055] (3) Add 200 μl BufferPP, vortex for 20 seconds, and incubate on ice for 3-5 minutes. Centrifuge at 13400×g for 3 minutes,...

Embodiment 3

[0080] 1. Primer design and preparation

[0081] Elferink et al. (2008) found that the slow feather K allele that controls feather speed is composed of tandem repeats with a total length of 176324bp, caused by partial duplication of two genes: PRLR gene and SPEF2 gene encoding sperm flagellin, including Exon 1-exon 11 and 558bp exon 12 of the PRLR gene and exon 1-exon 5 of the SPEF2 gene were identified; and primers were designed to amplify a 78bp long The breakpoint connects the sequence, but the fast feather chicken cannot amplify this sequence, so it can be used as a molecular detection method to distinguish fast and slow feathers. On the basis of the research results, primers were designed to amplify a breakpoint junction sequence containing 78bp and a length of 1305bp, which can only be amplified by slow-feathered chickens, but not by fast-feathered chickens, so that chickens can be distinguished The fast and slow feather type. The fragment is easy to amplify and has st...

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Abstract

The invention relates to the field of molecular genetics, and particularly provides molecular identification primers for chicken fast / slow feathering phenotype. The forward primer is 1305s: 5'CCTCCTTGCCAAACCTTA3'; and the reverse primer is 1305a: 5'ATCAGCCAGATCCGTCAG3'. The primers have the advantages of favorable specificity and high amplification efficiency, and can be used for molecular identification of chicken fast / slow feathering phenotype. The invention also provides a molecular identification method of chicken fast / slow feathering phenotype. When being used for experimentation in combination with the primers, the method is simple to operate and high in feasibility, and can solve the problems of undefined identification standard, low identification result accuracy, long male / female complete set line construction time and limited identification time in the fast / slow feathering identification.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a molecular identification primer and an identification method for chicken fast and slow feather phenotypes. Background technique [0002] Male and female identification is an important technology in modern chicken production. Since chickens are ZW-type sex-determined, the fast-slow feather gene locus is located on the Z chromosome, and the slow-feather is a dominant gene. Therefore, many chicken farms currently breed pure slow-feather lines through screening. And the pure fast-feather population, through the slow-feather hen and the fast-feather rooster, the commercial generation chicks are separated from each other by fast-slow feathers, the fast-feather chicks are hens, and the cocks are slow-feather. The distinction between fast and slow feathers is mainly determined by the length of the main wing feathers and overlying main wing feathers on the wings of newborn chicks. [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 李兰会张秀玲李祥龙臧素敏张乐超刘春杨周荣艳
Owner HEBEI AGRICULTURAL UNIV.
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