Primer combination and kit for simultaneous detection of 93 bovine genetic defect genes and lethal haplotypes

Abstract

The invention discloses a primer combination and a kit for simultaneous detection of 93 bovine genetic defect genes and lethal haplotypes. Through use of the primer combination and the kit, casual mutation sites (SNP and insertion or deletion of short fragments) of the 93 genetic defect genes (such as PIRM syndrome, crooked tail syndrome, epidermolysis bullosa and the like) and the lethal haplotypes (HH1, HH3, HH4, HH5 and JH1) can be detected in one time, cattle with monogenic inheritance defect gene carriers and lethal haplotype carriers can be screened, and accurate guidance is provided for cattle breeding and genetic evaluation, herd selection and assortative mating, population genetic improvement, cultivation of new varieties and genetic resource protection. The primer combination and the kit have the characteristics of high throughput, low cost, high accuracy, and the like, is suitable for the varieties of beef cattle and dairy cattle, and can be widely used in the breeding and reproduction field of the cattle.

Description

technical field [0001] The invention relates to the technical field of animal husbandry, and relates to a detection method for determining whether cattle carry 93 bovine genetic defect genes and lethal haplotypes, specifically, using DNA extraction, PCR multiple amplification technology, single base Extended technology and flight mass spectrometry technology, genotyping 93 genetic defect genes and lethal haplotypes at the same time, to determine whether the cattle carry recessive genetic defect genes and lethal haplotypes, involving recessive genetic defect genes and lethal haplotypes in cattle Type elimination and individual labeling, selection and matching of cattle herds, screening methods for dairy cows with specific type of β-casein milk, etc. Background technique [0002] So far, there are 506 known genetic defects in cattle, 233 of which belong to Mendelian inheritance, of which 133 are known to cause mutations (http: / / omia.angis.org.au / home / ), most Mutations are har...

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Problems solved by technology

[0009] China authorized invention patent, CN200610150317.9, a method for detecting harmful genes of bovine CVM, the invention provides a method for detecting bovine vertebral deformity syndrome (CVM) carriers, the method first designs synthetic primers for bovine DNA samples Carry out polymerase chain reaction (PCR), then carry out single-strand conformation polymorphism (SSCP) analysis to the PCR product, judge whether the cattle carry CVM harmful gene according to the result of polyacrylamide gel electrophoresis, its shortcoming is that can only detect Single CVM carrying case, and poor repeatability

[0010] China authorized invention patent, CN201310617444.5, a primer composition for the detection of harmful genes of tauridylate synthase deficiency and its kit and application, the invention discloses a detection of harmful genes of tauridin synthase deficiency The primer composition and kit and application thereof, the detection method of the invention is to first extract the whole group of DNA in bovine blood as a template, carry out nested PCR (NestedPCR) amplification, and the obtained PCR products are sequenced, and according to the sequenced results can be Directly understand the base change of the mutation site, which is characterized by two amplifications and sequencing of the product, which is easy to introduce errors and can only detect a single defective gene

[0011] China authorized invention patent, CN200910272954.7, invented a method for detecting tauridylate synthase deficiency by using AS-PCR. The method of enzyme deficiency, prepared a specific PCR primer for the detection method of tauridylate synthase deficiency, introduced mismatched bases in the amplified product, and needed to use electrophoresis for genotyping, because the product The length is short, it is easy to cause misjudgment, and it is not suitable for high-throughput and large-scale herd detection

[0012] The Chinese authorized invention patent CN201110440150.0 discloses a method for screening bovine degenerative axon disease carriers and its kit. The invention uses the genomic DNA of the cattle to be tested as a template, and designs upstream primers and downstream primers to carry it out. PCR amplification reaction, and then recover the PCR amplified fragment, digest it with FastDigest restriction enzyme TspRI, and perform gel detection after digestion in the enzyme digestion reaction system. This screening method is simple, fast, and low in cost, but there are also limitations Can detect the deficiency of a single genetic defect gene

[0013] At present, there is no research on methods for detecting other genetic defect genes and lethal haplotypes at home and abroad, and there is no research on high-throughput detection methods that can simultaneously detect hundreds of known genetic defect genes in cattle.

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