应用巢式PCR法检测猪单个胚胎中的基因突变

doi:10.16431/j.cnki.1671-7236.2016.10.009

Abstract

Abstract:

Gene mutation detection in single cell is a new method used for DNA or RNA analysis based on single cell lysis products,the establishment of this technology will provide an easy and quick way for gene expression and mutation detection in single cell level.In this paper,pig 4-cell-stage parthenogenetic embryos microinjected with Cas9 mRNA and 2 target gRNA from pig SS gene were used as experimental material,and two pairs of nested primers were designed for nested PCR.Using this detection method,long fragment deletion in SS gene was detected.This method can minimize the occurrence of false negative and false positive results,which may play an important role in the pig gene mutation detection.

Key words: nested PCR; single cell lysis; CRISPR/Cas9 system; gene mutation

CLC Number:

S814.6

REN Hong-yan, Xiao Hong-wei, LI Xiao-min, XU Zhen-yu, BI Yan-zhen, HUA Zai-dong, ZHENG Xin-min. Establishment and Application of Nested PCR Method for Detecting Mutation in Single Cell[J]. , 2016, 43(10): 2566-2571.

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Establishment and Application of Nested PCR Method for Detecting Mutation in Single Cell

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