Single nucleotide polymorphism (SNP) classification method for identifying inbred line mice

A typing method and inbred line technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of unsuitable genome research, reduce the number of reactions and time, save costs, and operate easily and quickly Effect

Inactive Publication Date: 2012-07-18
DONGHUA UNIV
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Problems solved by technology

[0002] The source and purity of experimental animals have an important impact on the experimental results, and a high-throughput genotyping platform for genetic quality monitoring and strain identification of inbred mice needs to be established urgently
There are many kinds of molecular genetic markers, but most of them are not suitable for large-scale genome research

Method used

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  • Single nucleotide polymorphism (SNP) classification method for identifying inbred line mice
  • Single nucleotide polymorphism (SNP) classification method for identifying inbred line mice
  • Single nucleotide polymorphism (SNP) classification method for identifying inbred line mice

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Obtain the sequence of selected SNP sites from NCBI (National Center for Biotechnology Information), and then design primers for these sites, and the main primer design is mainly completed by the Primer3 program.

[0019] Primer and probe sequences are listed in Table 1 and Table 2.

[0020] Table 1 Genome PCR amplification primer sequence

[0021]

[0022]

[0023]

[0024] Table 2 Genome LDR probe sequence

[0025]

[0026]

[0027]

[0028]

[0029]

[0030] (2) PCR-LDR scheme

[0031] During the multiplex PCR-LDR reaction, the annealing temperature is one of the main parameters, and choosing different annealing temperatures has a great influence on the connection efficiency. Analyzing the results of ABI377 sequencing electrophoresis, the optimal annealing temperature was finally determined to be 56°C.

[0032] The PCR reaction system is shown in Table 3:

[0033] Table 3 PCR reaction system

[0034]

[0035] The PCR reaction proce...

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Abstract

The invention relates to a single nucleotide polymorphism (SNP) classification method for identifying inbred line mice, which comprises (1) genetic mark choose, evenly covering 45 SNP locus of 21 chromosomes and designing primers and probes for the locus; (2) four groups of multiple polymerase chain reaction (PCR) and each group of the PCR containing 10 to 12 pairs of specificity probes of the locus; (3) four groups of multiple ligase detection reaction (LDR) and each group of the LDR containing 10 to 12 pairs of locus specificity probes; and (4) fast separation and identification of products. The SNP classification scheme only needs 45 SNP genetic locus to be used for classifying and identifying the inbred line mice, thereby reducing reaction times and time, being convenient and quick in operation, greatly saving cost and having significance on classification and researches. The SNP classification method is a new method for identification and classification of inbred mice.

Description

technical field [0001] The invention belongs to the field of strain identification and genetic quality detection of inbred mice, and in particular relates to a SNP typing method for identifying inbred mice. Background technique [0002] The source and purity of experimental animals have an important impact on the experimental results, and a high-throughput genotyping platform for genetic quality monitoring and strain identification of inbred mice needs to be established urgently. There are many kinds of molecular genetic markers, but most of them are not suitable for large-scale genome research. A large amount of SNP information has been found through the comparison of the complete genome sequence of C57BL / 6 and other inbred mouse strains. The number of SNPs is huge and can reduce the cost of genotyping. Tens of thousands of SNPs have been found among mouse strains and used as genetic markers for genotyping and QTL analysis of mice. Duality genetic marker single nucleotide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李凯谢雯鲍世民谢建云姜宪环蔡慧强
Owner DONGHUA UNIV
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