Primer composition for detecting harmful gene of deficiency of uridine monophosphate synthase of cattle, kit with primer composition and application of kit

Abstract

The invention discloses a primer composition for detecting a harmful gene of deficiency of uridine monophosphate synthase of cattle, a kit with the primer composition and an application of the kit. The primer composition disclosed by the invention is composed of a primer group A and a primer group B, wherein the primer group A is composed of a primer 1 and a primer 2, the primer group B is composed of a primer 3 and a primer 4, and the nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively shown as SEQIDNO. 1-4. The invention also provides the kit with the primer composition. The method for applying the kit disclosed by the invention to the detection of the harmful gene of the deficiency of uridine monophosphate synthase of cattle comprises the steps of extracting the complete set of DNA (Deoxyribonucleic acid) in cattle blood as a template to carry out nested PCR (Polymerase Chain Reaction) amplification to obtain a PCR product, and sequencing the obtained PCR product so as to directly know about the basic group change on a mutation site according to a sequenced result, thereby ensuring the accuracy of the result and meeting the requirements of a detecting technology for characteristics such as high speed, precision, high throughput and the like.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, it relates to a primer composition for detection of harmful genes of tauridylate synthase deficiency, a kit and application thereof. Background technique [0002] With the popularization and application of artificial insemination and embryo transfer technology, the wide application of dairy cow germplasm (embryo, semen, and bulls) around the world has significantly improved the genetic performance and accelerated the impact of recessive genetic diseases on cattle. group hazards. In particular, an excellent breeding bull can produce hundreds of thousands of doses or even millions of doses of frozen semen in its lifetime. If it carries a recessive genetic defect gene, it may spread rapidly all over the world and bring huge economic benefits to the production of animal husbandry. loss. my country mainly imports a large number of excellent germplasm of dairy cows from the Un...

AI Technical Summary

Problems solved by technology

The PCR-RFLP method is to design an AvaI enzyme cutting site at the mutation point, digest the PCR product, and judge whether there is an AvaI enzyme cutting site according to the enzyme digestion result. However, due to the large genome of cattle, PCR is only performed once. Due to the relationship between the concentration and purity of the DNA template, the amplification efficiency is not high, and error fragments are prone to occur, and there is a possibility of false positives. After the method introduces mismatched bases, the amplification efficiency of PCR will be affected

The AS-PCR method designs specific primers according to the mutation site, one of which is complementary to a base state of the mutation site, and the other chain is designed according to a conventional method. The specific primer has an amplified product in one genotype, There is no amplification product in another genotype, and it is determined whether there is a base mutation according to the presence or absence of the amplification product, but this method sometimes causes the base at the 3' end of the primer to be non-complementary to the base of the template In some cases, the extension can still be carried out, which may easily lead to misjudgment of the result

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