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

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

Article DOI: 10.3791/58467-v • 08:01 min • November 20th, 2018
November 20th, 2018 •
*1,2, *1, *3, 1, 1, 1, 4, 5, 3, 1
1Lab of Animal Models and Functional Genomics (LAMFG), The Key Laboratory of Animal Vaccine & Protein Engineering, College of Veterinary Medicine, Hunan Agricultural University (HUNAU), 2Department of Pathology, Georgetown University Medical School, 3College of Food Science and Technology, Hunan Agricultural University (HUNAU), 4Lab of Molecular Cardiology (LMC), National Heart, Lung, and Blood Institute (NHLBI)/National Institutes of Health (NIH), 5Transgenic Core, National Heart, Lung, and Blood Institute (NHLBI)/National Institutes of Health (NIH)
* These authors contributed equally

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Here, we present a detailed protocol for identifying homologous recombination events that occurred in mouse embryonic stem cells using Southern blotting and/or PCR. This method is exemplified by the generation of nonmuscle myosin II genetic replacement mouse models using traditional embryonic stem cell-based homologous recombination-mediated targeting technology.

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Homologous Recombination Events Mouse Embryonic Stem Cells Southern Blotting Polymerase Chain Reaction Molecular Biology Cellular Biology Accurate Technique Reliable Technique Genetically Modified Cells Genetically Modified Animals Multiple Steps GDNA Digestion Agarose Electrophoresis Gel Ethidium Bromide DNA Denaturing Solution DNA Neutralizing Solution
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