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Chinese hamster microsatellite genetic marker and screening method thereof

A technology of microsatellite marker and genetic marker, applied in the field of Chinese hamster microsatellite genetic marker and its screening

Inactive Publication Date: 2008-12-24
SHANXI MEDICAL UNIV
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AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems that there are no reports about Chinese hamster microsatellite markers, and most of the existing methods for screening microsatellite markers use enzyme digestion to break the DNA genome of species, the present invention provides a Chinese hamster microsatellite marker. Microsatellite Genetic Markers in Hamsters and Their Screening Methods

Method used

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  • Chinese hamster microsatellite genetic marker and screening method thereof
  • Chinese hamster microsatellite genetic marker and screening method thereof
  • Chinese hamster microsatellite genetic marker and screening method thereof

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Experimental program
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Embodiment Construction

[0011] 1. Construction of Chinese hamster genome microsatellite enrichment library

[0012] 1. DNA extraction

[0013] (1) Take Chinese hamster tail tissue, cut it into pieces with scissors, add 500 μl of lysis buffer, mix well, add proteinase K to a final concentration of 100L, mix well, and digest overnight at 50°C on a slow shaker.

[0014] (2) Cool the solution to room temperature, add an equal volume of Tris saturated phenol, and slowly invert until the two phases are mixed to form an emulsion. Centrifuge at 13200 / rpm at 4°C for 10 minutes, and slowly transfer the supernatant to another centrifuge tube with a 1ml pipette tip.

[0015] (3) Add an equal volume of Tris saturated phenol and chloroform, shake well, centrifuge at 13200 / rpm for 10 min at 4°C, and transfer the supernatant to another centrifuge tube.

[0016] (4) Add an equal volume of chloroform, shake well, centrifuge at 13200 / rpm for 10 min at 4°C, and transfer the supernatant to another centrifuge tube.

[...

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Abstract

The invention relates to an animal DNA molecule heredity mark technique, in particular to a Chinese hamster microsatellite heredity mark and a selecting method thereof to solve the problems that the report on the Chinese hamster microsatellite mark is not presented and the method for selecting the microsatellite mark that the DNA genome is broken down by the restriction enzyme digestion has a plurality of problems. The Chinese hamster genome DNA is extracted and is broken down by ultrasonic, and the DNA segment is reclaimed by electrophoresis. A Chinese hamster genome microsatellite enriched library is established, positive cloning vectors are selected from the library for sequencing. 17 microsatellite marks are obtained by selection. According to the microsatellite repetitive sequence, a primer is designed and 17 pairs of primers are selected for the heredity detection. When the method is used, the PCR amplification is performed on genome DNA of different individuals in a colony or of different colonies of Chinese hamster by using the 17 pairs of the primers. The electrophoresis detection is performed on the amplification product. According to the detection data and a map, the gene of each individual is determined. The method has good stability, high repetitiveness, strong comparability and simple operation.

Description

technical field [0001] The invention relates to an animal DNA molecular genetic marker technology, in particular to a Chinese hamster microsatellite genetic marker and a screening method thereof. Background technique [0002] Microsatellite (Microsatellite), also known as simple sequence repeats (Simple sequence repeats, SSR), refers to a simple multiple tandem repeat sequence composed of a few nucleotides (generally 1-6) through multiple repeat units, such as (CA) n , (TG) n etc., also known as short tandem repeats, simple sequence repeats are mostly within 100bp in length, and are widely distributed in eukaryotic genomes. There are conserved DNA sequences on both sides of the microsatellite sequence. According to the primers involved in the conserved sequences on both sides, the sequence of the microsatellite site can be amplified by PCR. Due to the different repeating times of the microsatellite repeating unit, the length of the amplified microsatellite sequence presen...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12
Inventor 宋国华胡松年刘田福耿佳宁岳文斌高莉
Owner SHANXI MEDICAL UNIV
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