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A method for monitoring the genetic quality of inbred mice using microsatellite technology

A technology of inbred strains and mice, applied in recombinant DNA technology, biochemical equipment and methods, microbial measurement/testing, etc., can solve the problems of low primer specificity and polymorphism, and easy judgment errors

Active Publication Date: 2022-04-19
上海斯莱克实验动物有限责任公司
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The number of microsatellite loci in the existing database is fixed, and the specificity and polymorphism of the corresponding primers recorded in the literature are not high; in addition, when judging the results of the loci of dibasic repeat sequences, judgment errors are prone to occur due to the influence of shadow bands
[0005] There are still many problems in the identification of inbred mice. It is still difficult to identify more strains of mice through simple and convenient methods in this field.

Method used

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  • A method for monitoring the genetic quality of inbred mice using microsatellite technology
  • A method for monitoring the genetic quality of inbred mice using microsatellite technology
  • A method for monitoring the genetic quality of inbred mice using microsatellite technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] Embodiment 1, the search of mouse microsatellite locus

[0128] 1. Preliminary screening

[0129] The whole genome sequence of mouse GRCm38.p2 was obtained from the GenBank mouse database. Afterwards, the inventors used BamHI and EcoRI to cut the genome sequence into short sequences, analyzed the short sequences with a length of 100-600 one by one, and searched for repeat sequences of 3, 4, 5, and 6 bases. The statistical results are shown in Table 1.

[0130] Table 1. Screening statistics of mouse microsatellite loci

[0131]

[0132] 2. Further screening

[0133] A search was performed on each chromosome in turn to pick a locus for a four-base repeat sequence containing "TATC". The inventors found that there are still very many such sites, which need to be further screened. In combination with experience and analysis, except the Y chromosome, the inventor selected suitable sites on each chromosome, which were not covered in the Genbank database, and initially ...

Embodiment 2

[0148] Embodiment 2, inbred mouse identification

[0149] In this example, use the primer pairs screened in the preceding Example 1 to identify inbred mice and determine whether different inbred mice can be distinguished: BALB / c, C57BL / 6, CBA, DBA / 1, DBA / 2, FVB, C3H / He, NOD, SJL / J.

[0150] Collect the mouse tails of 9 mouse strains, namely BALB / c, C57BL / 6, CBA, DBA / 1, DBA / 2, FVB, C3H / He, NOD, SJL / J, and extract their genomic DNA respectively , DNA concentration was adjusted to 50ng / ul.

[0151] PCR reaction system total 15ul):

[0152]

[0153]

[0154] PCR reaction program:

[0155] First, pre-denature at 94°C for 3 minutes;

[0156] After that, denaturation at 94°C for 15s, annealing at 55°C for 15s, extension at 72°C for 30s, 35 cycles.

[0157] ABI3730 sequencer electrophoresis:

[0158] Mix 1.5 ul of the 10-fold diluted PCR product with 0.25 ul of the ROX500 internal standard, incubate at 95°C for 5 min, and then rapidly ice-bath for 3 min. An ABI3730 seque...

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Abstract

The invention relates to a method for monitoring the genetic quality of inbred mice by using microsatellite technology. The present invention provides a series of microsatellite loci with good polymorphism and good polymorphism distributed on different chromosomes of mice and their specific primers, which are used for the identification and inheritance of commonly used inbred mouse strains Polymorphism monitoring. The method of the invention is simple and fast in operation, low in cost and easy to popularize and use.

Description

technical field [0001] The invention belongs to the field of species monitoring, and more specifically, the invention relates to a method for monitoring genetic quality of inbred mice by using microsatellite technology. Background technique [0002] The genetic background and response characteristics of experimental animals are important factors affecting the experimental results. Genetic variation, or differences between different sub-strains, will cause differences in experimental results, or even erroneous results. In addition, operational management errors in the breeding and breeding of experimental animals can also cause changes in the genetic background of experimental animals. With the development of biomedicine, the requirements for the quality of experimental animals are getting higher and higher, so the genetic monitoring methods for the quality of experimental animals also put forward higher requirements. In the process of production, reproduction or scientific ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 韩琳鲍世民
Owner 上海斯莱克实验动物有限责任公司
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