Method for identifying single-parent water buffalo parental right, primer and reagent kit thereof

A technology of paternity relationship and identification sheet, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of group inbreeding, population degradation, etc., achieve high accuracy, low sample requirements, and improve stability Effect

Inactive Publication Date: 2009-08-19
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Abstract
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Problems solved by technology

Errors caused by pedigree information errors will be magnified step by step throughout the breeding process, and it is very likely to cause large-scale inbreeding of the population, resulting in population deg...

Method used

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  • Method for identifying single-parent water buffalo parental right, primer and reagent kit thereof
  • Method for identifying single-parent water buffalo parental right, primer and reagent kit thereof
  • Method for identifying single-parent water buffalo parental right, primer and reagent kit thereof

Examples

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Effect test

Embodiment 1

[0051] Example 1. Screening of microsatellite markers and mitochondrial primers for paternity identification of Chinese buffaloes

[0052] 1. Primer design

[0053] Referring to GenBank, after repeated screening tests, polymorphic-rich microsatellite primers were screened from the common bovine microsatellite markers recommended by the Roslin Institute in the United Kingdom. The sequences of the screened microsatellite primers are shown in Table 1.

[0054] The primers were designed according to the D-Loop region of common bovine mtDNA, and the internal primers were designed for the convenience of sequencing. The primer sequences are shown in Table 2. The primers were synthesized by TaKaRabao Bioengineering (Dalian) Co., Ltd. The length of the amplified fragment is expected to be about 900bp.

[0055] Table 1 Selected 8 microsatellite loci of STR loci and their primer sequences

[0056]

[0057] Table 2 Primer sequences of the selected mtDNA D-Loop region

[0058]

[0...

Embodiment 2

[0107] Embodiment 2, carry out paternity identification to Chinese buffalo with microsatellite marker

[0108] The method for identifying the paternity relationship of single-parent buffalo in this embodiment comprises the following steps: screen polymorphic-rich microsatellite primers from common bovine microsatellite markers; Under the guidance of microsatellite primers, the blood samples of a cow (m), a Chinese buffalo known to be the offspring c1 of the cow and another suspected c2 of the cow's offspring were collected from a farmer's house in Xichang, Sichuan, China Carry out PCR amplification; conduct 8% polyacrylamide gel electrophoresis detection on microsatellite PCR amplification fragments; judge the genotype of each individual according to the electrophoresis detection results, and calculate the uniparental parentage index PI value and parent-child relative probability value according to the genotype W value, according to the W value to judge whether the parental re...

Embodiment 3

[0127] Example 3 Using Microsatellite Markers and Mitochondrial Genetic Markers to Identify the Parentage of Chinese Buffaloes

[0128] The microsatellite marker paternity testing method is the same as in Example 2.

[0129] The paternity identification of Chinese buffalo by mitochondrial genetic markers includes the following steps: design primers according to the D-Loop region of common bovine mtDNA, and the primer sequences are shown in Table 2; A farmer in Xichang, Sichuan, China collected blood samples from a cow, a buffalo known to be the offspring of the cow, and another buffalo suspected to be the offspring of the cow, and performed PCR amplification in a 50 μL reaction system, and the PCR amplification fragment of mitochondria Agarose gel purification was carried out, and the purified PCR products were sequenced by the dideoxy termination method, and the sequencing results were analyzed and judged by DNAman software.

[0130] 1. PCR amplification

[0131] According ...

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Abstract

The invention discloses a method for identifying single-parent buffalo paternity relationship as well as primers and kits thereof, belonging to the technical filed of genetic engineering. The method comprising the steps: screening out eight microsatellite primers with rich polymorphism; PCR amplifying by taking detected buffalo genome DNA as a template; detecting PCR amplified fragment through polyacrylamide gel electrophoresis; judging individual genotype according to the detected result; counting single-parent paternity index PI value and parent-child relative probability value W value according to the genotype; and judging whether the paternity relationship is existed or not according to the W value. In order to further improve identification stability, accuracy and sensitivity, the method designs primers so as to identify mitochondrial inheritance according to the D-Loop region of common cattle mtDNA. The method combines microsatellite method with mitochondrial inheritance marker method, and identifies from different angles and reaches consistent result, thereby improving identification stability, accuracy and sensitivity and having the advantages of wider identifying range and lower sample requirement.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for paternity identification of Chinese buffalo using microsatellite and mitochondrial genetic marker technology, special primers and a kit. Background technique [0002] In modern animal production and breeding, it is necessary to use the phenotypic information of various relatives to determine accurate genetic pedigrees and understand the breeding value of male animals through correct pedigrees, so as to determine individual selection and avoid individual inbreeding. With the development of large-scale joint breeding and the wide application of BLUP technology, the influence of pedigree information on breeding is increasing. Errors caused by pedigree information errors will be magnified step by step throughout the breeding process, and it is very likely to cause large-scale inbreeding of the population, resulting in population degradation, resulting in irr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 徐亚欧孙蕊毛亮
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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