Rapid visual detection method for sheep multi-fetus major gene FecB

Abstract

The invention discloses a rapid visual detection method for a sheep multi-fetus major gene FecB. The detection method comprises the following steps: (1) collecting a muscle tissue sample of a sheep individual and extracting genome DNA of the muscle tissue sample; (2) searching the DNA sequence of the FecB gene from a GeneBank database, and designing an ARMS specific primer aiming at the SNP site of the FecB, wherein the last base at the 3' end of a downstream primer is complementary with the SNP site, and the length of the primer is about 20bp; and (3) adding the nucleic acid dye SYBR Green Iinto a PCR system after the reaction by adopting the primers, and judging whether the sheep carries the multi-fetus major gene FecB or not by directly observing the color change of the reaction system, wherein the sample carrying the FecB is bright green, otherwise, the sample carrying the FecB is orange yellow. The detection method disclosed by the invention is high in sensitivity and accuracy, and can be used for rapidly and visually detecting the multi-fetus major gene of the sheep at low cost.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a rapid visual detection technology of FecB, the main effect gene for multiple births in sheep. Background technique [0002] Sheep play an important role in both agricultural production and people's life, and are very important livestock animals. Fertility and reproductive efficiency are key factors affecting the economy of sheep farming (Wang W, La Y, Zhou X, et al. The genetic polymorphisms of TGFβ superfamily genes are associated with litter size in a Chinese indigenous sheep breed(Hu sheep)[J]. Animal reproduction science, 2018, 189:19-29.), so the multiparity of sheep is the most important economic trait in the sheep breeding industry. Relevant studies have shown that the role of the main effect gene of multifetal performance can significantly increase the yield of mutton. However, since the vast majority of sheep breeds produce single lambs and f...

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Problems solved by technology

[0003] There are many methods for detecting FecB gene, such as PCR-RFLP method is widely used to analyze the mutation of FecB gene (Georgescu S E, Hrinca G, Rebedea M, et al. Investigation of FecBMutation in Four Romanian Sheep Breeds[J].Scientific Papers Animal Science and Biotechnologies, 2011,44(1):219-222.), but the primers of this method need to have enzyme cutting sites, and need to use expensive restriction enzymes, the method is cumbersome to operate and the detection cost is high

Other methods such as DNA sequencing or PCR-SSCP, these methods have disadvantages in terms of accuracy, cost, resolution, labor and time consumption, which become unacceptable as the sample size increases

The former, as the gold standard, not only requires a precise temperature-controlled circulation system, but also has high costs when the number of samples is large, and is not suitable for on-site detection; as for PCR-SSCP, the method is cumbersome to operate and has insufficient labor and time.

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