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Genetic markers and their applications for constructing and identifying molecular pedigrees of dairy cows

A technology of genetic markers and molecular genealogy, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of using a large number of microsatellite sites, cumbersome operation methods, and no reference value , achieving good application prospects and economic benefits, high detection efficiency, and optimized detection costs

Active Publication Date: 2019-08-09
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are microsatellite paternity identification methods and corresponding molecular markers for Holstein dairy cows. Although some technologies have disclosed some relevant content, due to differences between breeds or groups, there is basically no reference value and the molecular markers disclosed However, there are still some disadvantages, such as the use of a large number of microsatellite sites, the operation method is relatively cumbersome, and the cost of use is relatively high, and it is not suitable for a small number of or individual pedigree records or missing individuals that are common in dairy farms in production practice paternity test

Method used

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  • Genetic markers and their applications for constructing and identifying molecular pedigrees of dairy cows
  • Genetic markers and their applications for constructing and identifying molecular pedigrees of dairy cows
  • Genetic markers and their applications for constructing and identifying molecular pedigrees of dairy cows

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Selection and determination of microsatellite sites

[0029] 1. The microsatellite marker provided by the present invention consists of 8 sites, namely TGLA227, TGLA122, BMC1207, BM103, INRA037, INRA134, BP7 and MB026; the basic information of the 8 sites is shown in Table 4 below:

[0030] Table 4 Basic information of loci

[0031]

[0032]

[0033] The nucleotide sequence of the above primer sequence is shown in SEQ NO.1-16;

[0034] 2. Microsatellite locus genotype detection

[0035] Using single-plex fluorescent PCR combined with ABI3730 automatic sequencer fluorescence electrophoresis technology, the genotypes of PCR products at 8 sites in 300 dairy cows and 9 bulls were detected. The fluorescently labeled PCR amplification products were subjected to capillary electrophoresis, and the genotype peak was detected by ABI3730 sequencer, and the microsatellite genotype was determined by Gene Mapper V3.0 software combined with Gene Marker V1.75 softwar...

Embodiment 2

[0064] Example 2 Pedigree Accuracy Verification

[0065] In order to verify the accuracy and reliability of 8 microsatellite loci for paternity testing in dairy cow populations, this example carried out paternity testing on 8 pairs of father-son cows with paper pedigree records and father-son relationships. Genomic DNA of 8 parent-child pair cattle and 1 unrelated father cattle were extracted, and the genotypes and gene frequencies of 8 microsatellite loci were analyzed. The following results are shown in Table 10. Use the Parentage analyzes is module of the Analyze is program in the Cervus3.0 software to carry out data analysis. The LOD value in the table is the logarithmic value of the paternity index (paternity index). If the LOD value is greater than 0, it means that compared with any individual, the candidate parent (Candidate Parent) is most likely to be the real parent; LOD value less than 0 means that compared with any individual, the candidate parent cannot be the rea...

Embodiment 3

[0072] Example 3 Application of 8 microsatellite loci in identification of genetic relationship in dairy cows

[0073] In this example, a pair of parent-offspring with a clear parent-child relationship and an unrelated parent were selected from the dairy cow population for verification, simulating parentage identification under normal production conditions.

[0074] sample

[0075] (1) 13325 blood samples of offspring cows, No. 1;

[0076] (2) Frozen semen of candidate male bull 37310001, No. 2;

[0077] (3) Frozen semen of irrelevant male bull 37310026, number 3.

[0078] Identification requirements: DNA test, paternity test

[0079] Test and Results

[0080] Extract the DNA of the above-mentioned sample to be tested, and perform PCR amplification on 8 microsatellite loci;

[0081] Taking the 15 μl amplification system as an example, the reaction system for PCR amplification is shown in Table 2.

[0082] Table 2 PCR amplification system

[0083]

[0084] The PCR rea...

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Abstract

The invention relates to molecular genetics, and provides genetic markers for establishing and identifying a dairy cattle molecule system spectrum and application of the genetic markers. The genetic markers consist of 8 loci which are respectively TGLA227, TLA122, BMC1207, BM103, INRA037, INRA134, BP7 and MB026. The invention further provides primers of the 8 microsatellite loci, a method for obtaining fluorescence labeling primers by using the primers to perform fluorescence labeling capillary electrophoresis, a method for performing genotype judgment by using Gene Mapper V3.0 software together with Gene Marker software V1.75, a method for performing genetic relationship identification by using Cervus3.0 software, and a method for establishing an application kit. The genetic markers can be applied to dairy cattle system spectrum identification and establishment, are capable of making important basis for increasing the accuracy of dairy cattle species selection and breeding, are capable of promoting the breeding progress of dairy cattle, and have good application prospects and economical benefits.

Description

technical field [0001] The invention relates to molecular genetics, and provides a group of genetic markers for constructing and identifying dairy cow molecular pedigrees and applications thereof. Background technique [0002] The genetic improvement of dairy cows requires the determination of the offspring of breeding bulls and molecular marker-assisted selection of dairy cows, and the accuracy of pedigree records is crucial to the genetic improvement of dairy cows. The lack of strict management of some small and medium-sized dairy farms leads to the lack of dairy cow pedigree, or due to the popularization of artificial insemination technology and inevitable errors in the production process (pedigree record), resulting in incomplete or wrong pedigree records of cow herds, so the molecular pedigree construction and pedigree of dairy cows Accuracy assessment has become an important part of genetic improvement. [0003] Microsatellite marker technology is a relatively mature ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 师科荣王中华殷彬岳书俭
Owner SHANDONG AGRICULTURAL UNIVERSITY
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