Polymorphic primers for Bactrian camels and their screening methods and methods for identifying parent-child relationship

A polymorphic primer, parent-child relationship technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as heavy workload and easy loss, and achieve high polymorphism, The effect of broad application prospects

Active Publication Date: 2017-03-01
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at a series of problems such as the current paternity identification method of Bactrian camels can only be recorded by traditional methods, the workload is heavy and easy to be lost, the present invention provides a polymorphic primer and Its screening method, and the method for identifying parent-child relationship with said polymorphic primer

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  • Polymorphic primers for Bactrian camels and their screening methods and methods for identifying parent-child relationship
  • Polymorphic primers for Bactrian camels and their screening methods and methods for identifying parent-child relationship
  • Polymorphic primers for Bactrian camels and their screening methods and methods for identifying parent-child relationship

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This embodiment provides a polymorphic primer for microsatellite molecular markers in the Bactrian camel transcriptome and a deletion method thereof.

[0041] 1. Polymorphic primer screening of Bactrian camel transcriptome microsatellite molecular markers

[0042] The transcriptome data of wild Bactrian camels were downloaded from the NCBI public database, and a total of 23,624 transcriptome sequences were obtained, which were independently stored in the FASTA format. Use MIcroSAtellite (MISA http: / / pgrc.ipk-gatersleben.de / misa / misa.html) to analyze the Bactrian camel transcriptome sequence, and screen the Bactrian camel microsatellite molecular sequence. The screening criteria are: repeats of two bases, three bases, four bases, five bases and six bases are respectively 7, 6, 5, 4, 3 and 2 (single nucleotide repeats are not selected because It is generally considered that it is not a useful polymorphic marker); a total of 1820 SSR sites were obtained, and the ratio of ...

Embodiment 2

[0063] The present embodiment provides a method for identifying the parent-child relationship of Bactrian camels using the polymorphic primers of the Bactrian camel transcriptome microsatellite molecular markers provided in Example 1, specifically as follows:

[0064] First use cervus 3.0 software to count the PE1 (non-parent exclusion rate: the genotypes of both parents are known), PE2 (non-father exclusion rate: only know the genotype of the father or mother) of the 117 Bactrian camel samples in Example 1 and the total non-parent exclusion probability (PE total ), 117 Bactrian camel samples PE1, PE2 and PE total See Table 5 for details. PE1, PE2, and PE total Calculation reference formula (Jamieson A. & Taylor S.C. Comparisons of three probability formula for parentage testing, Animal Genetics, 1997, 28, 397-400):

[0065] Paternity Exclusion Rate (PE1):

[0066]

[0067] Paternity Exclusion Rate (PE2):

[0068]

[0069] The overall paternal exclusion rate (PE to...

Embodiment 3

[0093] This embodiment provides a polymorphic primer of 14 pairs of Bactrian camel transcriptome microsatellite molecular markers provided in Example 1 to analyze the genetic diversity of Bactrian camels for revealing the genetic diversity of Bactrian camel samples Current situation, based on 14 pairs of polymorphic primers of microsatellite molecular markers in the Bactrian camel transcriptome and the diversity information detected in 117 Bactrian camel samples, the diversity analysis of each population was performed using PopGene1.32 software, in which:

[0094] Calculate the main genetic diversity parameters and observe heterozygosity (observed heterozygosity, Ho);

[0095] Expected heterozygosity (expected heterozygosity, He);

[0096] Number of alleles (number of alleles, N A );

[0097] Effective number of alleles (Ne);

[0098] Polymorphism information content (polymorphism information index, PIC) calculation reference formula (BotsteinD, White RL, Skolnick M, Davis ...

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Abstract

The invention discloses a polymorphism primer design with a two-humped camel transcriptome microsatellite molecular marker, a screening method of the polymorphism primer, and a method for identifying the parenthood of two-humped camels. The method for identifying the parenthood of two-humped camels by using the polymorphism primer comprises the following steps: acquiring two-humped camel whole blood samples with parenthood by utilizing an EDTA anticoagulant tube, and extracting the two-humped camel genome DNA by adopting a nucleic acid extraction kit method (a magnetic bead method); mixing the polymorphism primer with the two-humped camel transcriptome microsatellite molecular marker and the genome DNA, and performing PCR amplification, thereby obtaining an amplification product; performing 3730XL-type instrument capillary electrophoresis on the PCR product so as to detect an SSR molecular marker; and performing parenthood analysis on the result.

Description

technical field [0001] The invention relates to a polymorphism primer of a Bactrian camel transcriptome microsatellite molecular marker, a screening method thereof and a method for identifying parent-child relationship, belonging to the technical field of animal molecular biology. Specifically, the present invention relates to a method of selecting 18 SSR (Simple Sequence Repeat, Simple Sequence Repeat, SSR for short) molecular markers from Bactrian camel transcriptome data, and synthesizing their polymorphic primers respectively, and said polymorphic primers The morphological primers were used in the identification of the parent-child relationship of Bactrian camels. Background technique [0002] Accurate and rapid paternity identification of mammals plays an important role in the verification of parentage of mammals, the protection of kinship, the identification of true and false parentage, and property rights disputes of parentage. At present, paternity testing experimen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 吉日木图明亮
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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