A rapid and visual detection method for fecb, the main gene for multiple births in sheep

Abstract

The invention discloses a rapid and visual detection method for sheep multifetal main gene FecB. The steps of the detection method are: (1) collecting individual sheep muscle tissue samples and extracting its genome DNA; (2) searching for the DNA of the FecB gene from a GeneBank database Sequence, design ARMS-specific primers for the SNP site of FecB, that is, the last base at the 3' end of the downstream primer is complementary to the SNP site, and the length of the primer should be about 20bp; (3) Using the above primers, the nucleic acid dye SYBR Green Ⅰ was added to the PCR reaction system, and it was judged whether the sheep carried the main multiple-fetal gene FecB by directly observing the color change of the reaction system, that is, the sample carrying FecB was bright green, otherwise it was orange-yellow. The detection method of the invention has high sensitivity and accuracy, and can quickly, visualize, and low-cost detect the main effect gene of multiple births in sheep.

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 of sheep multiple births. Background technique [0002] Sheep play an important role in agricultural production and people's lives, and are very important livestock animals. Fertility and reproductive efficiency are the key factors affecting the sheep farming economy (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 sheep farming. Relevant studies have shown that the role of the major gene for multiple birth performance can significantly increase the yield of mutton. However, since the vast majority of sheep breeds produce single lambs and a small number of dou...

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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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