Mouse-tail DNA (deoxyribose nucleic acid) extraction kit applicable to the genotype of laboratory mouse and application thereof

Abstract

The invention discloses a simple, economic and reliable mouse-tail DNA (deoxyribose nucleic acid) extraction kit, which is applicable to accurate identification of the genotype of a laboratory mouse. The kit comprises liquid A and liquid B, wherein the liquid A consists of NaOH solution with the final concentration being 0.025N and EDTA (ethylene diamine tetraacetic acid) solution with the final concentration being 2mM; and the liquid B is Tris solution with the final concentration being 40mM and pH being 7.6. The application of the kit comprises the following steps of: taking 0.2-0.5cm tissue of the tail tip of the mouse, and putting the tail tip into a 1.5ml EP (eppendorf) tube; adding 180muml of liquid A, and putting the EP tube in a trace thermostat with the temperature being 100 DEG C for 30 minutes; and then carrying out ice bath for 2 minutes, then adding 180mul of liquid B, fully mixing to be uniform, and obtaining the tissue DNA which can be used for genotype identification in the next step. In the kit, the preparation method is simple, the components of the liquid A and the liquid B are conventional biochemical reagents respectively, the whole process does not need the reagents such as protease K, phenol / chloroform and the like to participate in and does not have fuzzy steps such as multiple centrifugation and the like as well, and the operation time is not more than1 hour, so that the invention has the advantages of fastness, economy and high cost performance, and is especially suitable for being used in laboratories for identifying the mouse genotype in a long-term, numerous and frequent manner.

Description

technical field [0001] The invention relates to a mouse tail DNA extraction kit and application thereof, which is simple, economical and reliable, and is especially suitable for accurate identification of experimental mouse genotypes. Background technique [0002] The use of genetics to study various gene mutant mice has become one of the most powerful methods in life science and medical research, and transgenic mice (mainly including gene knock-in, knock-out and immunodeficiency mice, etc.) The strains and numbers are also increasing. This phenomenon not only effectively improves the scientific research platform and level, but also makes the genotyping of mice a long-term, large-scale and frequent routine work. [0003] At present, the commonly used method involving tissue DNA extraction in genotyping is the extraction of rat tail DNA, and its traditional methods include phenol / chloroform extraction and high-salt extraction. Although these methods can obtain mouse tail DN...

AI Technical Summary

Problems solved by technology

Although these methods can obtain mouse tail DNA with higher purity and yield, they all have some common shortcomings: one is that the steps are more complicated, such as involving centrifugation many times; the other is that there are too many reagents involved, such as using Chloroform, ethanol, proteinase K and other reagents; the third is that the time is too long, for example, there is an overnight experiment step

The above characteristics have virtually increased the cost of experiments (including reagent costs and labor costs), which is not conducive to laboratories with tight funds and limited manpower to carry out related genotyping experiments

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