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Method and primer set for monitoring genetic quality of DBA inbred strain mice and applications thereof

An inbred line and primer set technology, applied in recombinant DNA technology, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of limited genetic profiles, low accuracy, and limited detection sites

Active Publication Date: 2019-04-30
GEMPHARMATECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using this method for detection has disadvantages such as low accuracy, low sensitivity, limited detection sites, and limited response genetic profile

Method used

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  • Method and primer set for monitoring genetic quality of DBA inbred strain mice and applications thereof
  • Method and primer set for monitoring genetic quality of DBA inbred strain mice and applications thereof
  • Method and primer set for monitoring genetic quality of DBA inbred strain mice and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] A method for monitoring the genetic quality of DBA inbred mice. It includes using primers to detect 96 SNP sites as shown in Table 5.

[0061] Table 5 96 SNP sites

[0062]

[0063]

[0064]

[0065] The sequences of primer combination 1 to primer combination 96 for detecting 96 SNP sites are specifically shown in Table 6.

[0066] Table 6 Sequences of primer combinations

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073] Specifically, the KASP method was used to perform PCR amplification on the treated sample with a primer set. The reaction system of the KASP method is: 0.8 μL of 25ng / μL DNA of the mouse to be tested, 0.778 μL of 2 × KASP Master Mix, and 0.022 μL of KASP Primer mix. Wherein, KASP Primer mix is ​​any combination of the above-mentioned primer combination 1 to primer combination 96.

[0074] During the detection process, DMSO with 5% volume of Master mix can be added, or Mg with 0.63% volume of Master mix can be ...

Embodiment 2

[0084] The method provided in Example 1 was tested on DBA inbred strain mice.

[0085] The DBA mice were typed using the method provided in Example 1, and IntelliQube fluorescence detection was used for reading, and IntelliScore was used for data analysis after PCR, and the genotype was automatically derived for analysis.

[0086] Using A / J Nju mice as a control strain, use the above 96 loci and primers to test the genotyping of DBA, please refer to Table 9.

[0087] Table 9A / J Nju is used as the DBA primer test result of control

[0088]

[0089]

[0090]

[0091] As can be seen from Table 9, the above 96 sites of DBA are all consistent with the site information registered on NCBI, and there are obvious differences between the specific sites and the control strain (A / J Nju mice). Therefore, the embodiments of the present invention 1 provides a method that can be used to monitor the genetic quality of DBA strain mice.

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Abstract

The invention relates to the fields of inbred strain mouse genetic background identification and genetic contamination detection, and provides a method and primer set for monitoring the genetic quality of DBA inbred strain mice and applications thereof. Specifically, the method comprises a step of detecting the genotype of specific SNP sites of a mouse to be detected so as to rapid identify the DBA strain from related inbred strains rapidly and efficiently; thus the genetic information of the DBA strain is effectively distinguished from the genetic information of other strains, the requirements of detecting the source of hybrid strains can be satisfied in genetic quality monitoring, and the method has the advantage that the cost for monitoring the genetic quality of DBA inbred strain miceis low.

Description

technical field [0001] The invention relates to the fields of genetic background identification and genetic pollution detection of inbred mouse strains, in particular, a method for monitoring the genetic quality of DBA inbred mice, a primer set and applications thereof. Background technique [0002] The quality control of experimental animals is the core issue for the healthy development of the experimental animal industry, and the quality control of mouse microorganisms and genetic background control are important control factors. [0003] Experimental animal strains refer to groups of animals with clear sources, which are bred by a certain mating method, have similar appearance, unique biological characteristics and stable genetic characteristics, and can be used for different experimental purposes. They have a clear genetic background and are strictly controlled. Genetically controlled animals. According to their genetic characteristics, they can be divided into inbred s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2535/125C12Q2563/107
Inventor 赵静琚存祥马秀英杨旭乐裴喆李永兵陶然张冬冬罗天同侯欢欢张明坤
Owner GEMPHARMATECH CO LTD
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