摘要
CRISPR-Cas9是一种现代生命科学研究中普遍使用的新型基因编辑技术。为了升级课程内容体系,使实验教学贴近科研实际,提升实验课程教学质量,设计了T7核酸内切酶1(T7E1)检测CRISPR-Cas9基因编辑效果实验。以CRISPR-Cas9基因编辑技术对mfn1编辑过的HeLa细胞为实验材料,提取基因组DNA,PCR扩增mfn1基因片段,切胶回收后,变性,退火,使用T7E1对编辑效果进行检测。经1%琼脂糖凝胶电泳,可观察到T7E1将略大于500 bp的mfn1基因片段切成两条大小约为250 bp的DNA片段,表明CRISPR-Cas9成功地对mfn1进行了编辑。
CRISPR-Cas9 is a new gene editing technology widely used in the research of modern life science.In order to upgrade the course content system,make the experimental teaching close to the scientific research practice,and improve the teaching quality of experimental courses,the experiment named testing the effect of CRISPR-Cas9 gene editing by T7 endonuclease 1(T7E1)is designed.This experiment used HeLa whose mfn1 has been edited by CRISPR-Cas9 as experimental materials,extracted the genomic DNA,amplified the mfn1 gene fragment by PCR,recovered the fragment by cutting gel,denatured,annealed and tested the editing effect by T7E1.Though the 1%agarose gel electrophoresis,it is observed that the mfn1 gene fragment slightly larger than 500 bp has been cut into two DNA fragments whose size is about 250 bp.This indicated that CRISPR-Cas9 has edited the mfn1 successfully.
作者
王宏刚
魏远
李艳君
朱玉山
WANG Honggang;WEI Yuan;LI Yanjun;ZHU Yushan(National Experimental Teaching Demonstration Center of Biology,Nankai University,Tianjin 300071,China)
出处
《实验技术与管理》
CAS
北大核心
2023年第5期38-41,共4页
Experimental Technology and Management
基金
2020年天津市高等学校本科教学质量与教学改革研究计划项目(B201005506)。
