中国畜牧兽医2016，Vol.43Issue(10)：2566-2571,6.DOI:10.16431/j.cnki.1671-7236.2016.10.009
应用巢式PCR法检测猪单个胚胎中的基因突变
Establishment and Application of Nested PCR Method for Detecting Mutation in Single Cell
摘要
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 impor-tant role in the pig gene mutation detection.关键词
巢式PCR/单细胞裂解/CRISPR/Cas9技术/基因突变Key words
nested PCR/single cell lysis/CRISPR/Cas9 system/gene mutation分类
农业科技引用本文复制引用
任红艳,肖红卫,李晓敏,徐振宇,毕延震,华再东,郑新民..应用巢式PCR法检测猪单个胚胎中的基因突变[J].中国畜牧兽医,2016,43(10):2566-2571,6.基金项目
转基因生物新品种培育重大专项(2016ZX08006001-005、2016ZX08006002-006、2016ZX08010003-006)；湖北省农业科技创新中心(2016-620-000-001-027)；湖北省科技支撑计划(2014BBB010)；湖北省农科院青年基金(2014NKYJJ02、2016NKYJJ19) （2016ZX08006001-005、2016ZX08006002-006、2016ZX08010003-006）
