Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

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期刊论文

作者：

Zhou, Dan;Tan, Lei;Li, Jian;Liu, Tanbin;Hu, Yi;...

通讯作者：

Wang, Aibing;Guo, Shiyin

作者机构：

[Hu, Yi; Liu, Tanbin; Wang, Aibing; Zhou, Dan; Tan, Lei; Li, Yalan] Hunan Agr Univ HUNAU, Coll Vet Med, LAMFG, Key Lab Anim Vaccine & Prot Engn, Changsha, Hunan, Peoples R China.

[Zhou, Dan] Georgetown Univ, Dept Pathol, Sch Med, Washington, DC 20057 USA.

[Guo, Shiyin; Li, Jian] Hunan Agr Univ HUNAU, Coll Food Sci & Technol, Changsha, Hunan, Peoples R China.

[Kawamoto, Sachiyo] NHLBI, LMC, NIH, Bldg 10, Bethesda, MD 20892 USA.

[Liu, Chengyu] NHLBI, Transgen Core, NIH, Bldg 10, Bethesda, MD 20892 USA.

通讯机构：

[Wang, Aibing; Guo, Shiyin] H

Hunan Agr Univ HUNAU, Coll Vet Med, LAMFG, Key Lab Anim Vaccine & Prot Engn, Changsha, Hunan, Peoples R China.

Hunan Agr Univ HUNAU, Coll Food Sci & Technol, Changsha, Hunan, Peoples R China.

语种：

英文

关键词：

Genetics;Issue 141;Homologous recombination;gene targeting;Southern blotting;PCR;mouse;embryonic stem cells;probe;primer;genetic replacement;knockout/knock-in;Myh9 gene;genomic DNA

期刊：

Journal of Visualized Experiments

ISSN：

1940-087X

年：

2018

卷：

2018

期：

141

DOI：

10.3791/58467

基金类别：

General Program of National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31571432]; Human Provincial Natural Science Foundation of China [2015JC3097]; Research Foundation of Education Bureau of Hunan Province, China [15K054]; NATIONAL HEART, LUNG, AND BLOOD INSTITUTEUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Heart Lung & Blood Institute (NHLBI) [ZICHL005907, ZICHL005907, ZICHL005907] Funding Source: NIH RePORTER

机构署名：

本校为第一且通讯机构

院系归属：

动物医学院

食品科学技术学院

摘要：

Relative to the issues of off-target effects and the difficulty of inserting a long DNA fragment in the application of designer nucleases for genome editing, embryonic stem (ES) cell-based gene-targeting technology does not have these shortcomings and is widely used to modify animal/ mouse genome ranging from large deletions/insertions to single nucleotide substitutions. Notably, identifying the relatively few homologous recombination (HR) events necessary to obtain desired ES clones is a key step, which demands accurate and reliable methods. S...

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Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

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