SNP marker related to tail type trait of fat tail type sheep, and applications thereof
A sheep and marker technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of prone to parasites, frostbite, anus, vulva exposure, etc., to promote the breeding process, save money time, low cost
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Embodiment 1
[0032] Example 1: Determination of a SNP marker related to the tail shape traits of fat-tailed sheep and its detection method and application
[0033] 1. Sample collection and DNA extraction
[0034] Randomly collect EDTA anticoagulant blood samples from 10 sheep in Ujimqin sheep, Sunit sheep, Barhu sheep, and Hulunbeier short-tailed sheep, and use a conventional blood genome extraction kit to extract genomic DNA from the blood;
[0035] 2. Specific primer sequence
[0036] According to the sequence of the T gene locus included in the NCBI database, primers for the screened T gene SNP site were designed, and the primer sequence was: forward primer (sequence listing sequence 2): 5'-CACAAGAAGGTGCAGAGTCACA-3'; reverse primer (sequence Listed sequence 3): 5'-CCTGCCTTTGGCCTCACGAT-3'.
[0037] 3. Amplify the target fragment in vitro
[0038] (1) Prepare 50μl PCR reaction system:
[0039] 50 ng / μl template DNA 4μl,
[0040] 10pmol / μl forward primer and reverse primer each 2μl, ...
Embodiment 2
[0059] Example 2: Kit and application for identifying tail-type traits of fat-tailed sheep
[0060] The components in the kit are as follows:
[0061] 10pmol / μl upstream primer 100μl
[0062] 10pmol / μl downstream primer 100μl
[0063] 2×Taq PCR MasterMix 1.25ml
[0064] Pas I endonuclease 50μl
[0065] 10×Buffer 100μl
[0066] Deionized water 2ml,
[0067] Total PCR reaction system in the kit:
[0068] Make up to 50μl with deionized water
[0069] 2×Taq PCR MasterMix 25μl
[0070] Template DNA 200ng
[0071] 10pmol / μl upstream primer 2μl
[0072] 10pmol / μl downstream primer 2μl;
[0073] PCR reaction program:
[0074] 94°C, 5 minutes; 94°C, 30 seconds; 57°C, 30 seconds; 72°C, 30 seconds; 35 cycles; 72°C, 10 minutes, 4°C storage.
[0075] Pas I digestion reaction system in the kit:
[0076] PasI endonuclease 1μl
[0077] 10×Buffer 2μl
[0078] PCR product 5μl
[0079] 12 μl of deionized water.
[0080] Pas I reaction procedure:
[0081] Reaction at 55°C for 6h...
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