Method utilizing fluorescence PCR technology to identify mouse genotype

Abstract

The invention relates to a method utilizing a fluorescence PCR technology to identify mouse genotype. A fluorescence PCR primer is utilized, after PCR amplification, the amplification product is mixed with ROX with a known size, the size of PCR products can be calculated by Genemapper software through 377 sequencing; and thus the mouse genotype can be identified. The provided method can simply, directly, and precisely find out the mice having micro RNA, whose Crispr / Cas9 has been knocked off.

Description

technical field [0001] The invention belongs to the field of mouse typing identification methods, in particular to a mouse typing identification method using fluorescent PCR technology. Background technique [0002] Since its inception, gene targeting technology has been one of the important means to study gene function, revealing the biological functions of many important genes. In addition, researchers also hope to use gene targeting technology to knock out or modify specific genes, so as to achieve the purpose of treating diseases or improving production traits of livestock and poultry. However, the efficiency of early gene targeting technology was extremely low, and it was difficult to truly apply it to medical treatment or livestock and poultry improvement. CRISPR / Cas9 system, as an emerging gene-directed editing technology, has gradually matured and been successfully applied in many animal and plant species, which has greatly promoted the study of gene function. The ...

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Problems solved by technology

However, this method is too cumbersome and needs to be annealed for a period of time before the next experiment can be carried out; using the enzyme digestion method, if the efficiency of the enzyme digestion is low, it will also produce false negative results; using agarose electrophoresis, the resolution is low

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