National Heart, Lung, and Blood Institute (NHLBI)/National Institutes of Health (NIH)

1 ARTICLES PUBLISHED IN JoVE

JoVE Journal

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

Dan Zhou^*1,2, Lei Tan^*1, Jian Li^*3, Tanbin Liu¹, Yi Hu¹, Yalan Li¹, Sachiyo Kawamoto⁴, Chengyu Liu⁵, Shiyin Guo³, Aibing Wang¹

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)

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.