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Genotypic assay kit for individual identification and paternity test of yaks

A technology for paternity testing and genotyping, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problems of failing to detect yak STR sites

Inactive Publication Date: 2018-03-13
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the STR marker primers of some common cattle can be used to amplify the corresponding loci of yaks, after all, there are certain differences in gene sequences between yaks and common cattle, and direct use of common cattle primers often cannot reach the yak STR loci. The best effect of detection

Method used

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  • Genotypic assay kit for individual identification and paternity test of yaks
  • Genotypic assay kit for individual identification and paternity test of yaks
  • Genotypic assay kit for individual identification and paternity test of yaks

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Screening and determination of STR markers for paternity identification of yaks:

[0097] 1. Selection and grouping of STR marker sites

[0098] 250 unrelated yaks were selected, and their venous blood DNA was extracted and tested. According to the relevant reports of literature research, 36 STR loci with no linkage relationship between each other and high polymorphic information content were selected, and the primers reported in the literature were sequenced with the corresponding loci in yak. Inappropriate primers, such as incompatible bodies and large differences in annealing temperature, were redesigned (see Table 7). In addition, four new STR loci (CH6AT, CH24AC, CH25GT, CHX4CA) were searched on the yak genome (see Table 7), and primers were designed for each STR locus and the sequences at both ends to amplify the corresponding loci. Detection of amplification products.

[0099] Table 7 Information of 40 pairs of yak STR common primers

[0100]

[0101]

...

Embodiment 2

[0125] A case of paternity identification of a yak triplet:

[0126] A pair of female yaks and offspring yaks with a clear mother-child relationship were selected from the yak parent-child group, and the offspring yaks had two suspicious fathers. The blood of the female yak, the offspring yak and two suspicious fathers (candidate father yak 1 and candidate father yak 2) were extracted respectively, and the parent-child relationship was tested with the kit of the present invention.

[0127] 1. Sample:

[0128] (1) The number of the offspring yak is B3916103, and the blood sample tube number is V111477;

[0129] (2) The number of the female yak is C0405066, and the blood sample tube number is Y72372390;

[0130] (3) The number of candidate male Yak 1 is C2010051, and the blood sample tube number is Y72372125;

[0131] (4) The number of the candidate male Yak 2 is C2310027, and the blood sample tube number is Y72372168.

[0132] 2. Extract the genomic DNA of 4 yak blood sampl...

Embodiment 3

[0142] A case of yak semen individual identification:

[0143] The two batches of preserved frozen semen are suspected to be collected from the same yak bull, and the blood sample of the yak bull and the two batches of frozen semen samples are taken, and the individual identification test is carried out with the kit of the present invention.

[0144] 1. Sample:

[0145] (1) The number of the yak bull is C0610069,

[0146] (2) The label of frozen semen 1 is 229137;

[0147] (3) The number of frozen semen 2 is 229138.

[0148] 2. Extract the genomic DNA of the yak bull blood sample and two batches of yak semen samples, and use the kit in the present invention to perform fluorescent primer amplification and capillary electrophoresis detection on each sample.

[0149] 3. Inspection and results

[0150] According to the difference in fluorescence signal of each amplification product in the above capillary electrophoresis, the length of the amplification product of the 15 STR si...

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Abstract

The invention discloses a composite PCR amplification system and an assay kit for genotyping of an STR mark detection system for individual identification and paternity test of yaks. The primer with afluorescence mark can achieve amplification of 15 STR mark sites of yaks, the mark sites are unlocked and have high heterozygosity and rich content of polymorphism information, and amplified productsare of right size. According to a detection method, genotyping results of 15 STR sites are obtained through amplification of 15 pairs of primers and electrophoresis detection; and individual identification and paternity test of yaks are carried out according to the STR locus genotyping results and gene frequency of each STR of yaks. Since individual identification and paternity test of yaks can be carried out effectively by comprehensively analyzing the STR locus genotyping results, accuracy of genealogy in yak breeding can be improved and the breeding process can be accelerated; accordingly,the composite PCR amplification system and the assay kit have good application prospect and economic value.

Description

technical field [0001] The invention relates to the technical field of in vitro detection of animal nucleic acid, in particular to a compound PCR amplification system and a detection kit for yak individual identification and paternity identification genotyping of a STR marker detection system. Background technique [0002] The pedigree information of livestock plays an important role in the breeding process. Incorrect pedigree will greatly reduce the accuracy of genetic evaluation, affect the effect of selection and mating, reduce the genetic progress of the population, and bring huge economic losses to animal production. In the production practice of yak breeding, errors in pedigree records are often unavoidable. It is affected by errors in breeding records, semen labeling, calving records, pedigree entry, and ear number loss. In grazing conditions, it is difficult to identify the father, or even the mother, of a calf after birth. Therefore, it is very necessary to carry o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2525/151C12Q2563/107C12Q2537/143C12Q2565/125
Inventor 裴杰郭宪褚敏包鹏甲王宏博丁学智吴晓云梁春年阎萍
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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