Molecular identification primers and method for chicken fast/slow feathering phenotype

[0040] Example 1

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

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

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

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

[0045] 4), analyze the electrophoresis results in 3).

[0046] Example 2

[0047] 1. Primer preparation

[0048] Primer synthesis was performed by Sangon Bioengineering (Shanghai) Co., Ltd., including:

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

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

[0051] 2. The steps of the kit to extract chicken blood clot DNA:

[0052] The ClotBloodDNAKit of 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 BufferGTL, 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

[0079] Example 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 a