Manipulation of mammalian genome and generation of genetically engineered mouse models (GEMMs) has revolutionized the scientific approach to address biological questions. To date, a number of gene-targeting strategies have been devised and are available to investigators for the generation of genetically modified mouse lines. Nevertheless, irrespective of the methodological approach selected, there remain critical molecular steps that need to be performed and put in place in order to obtain controlled and well-characterized new animal models. Here we provide technical details for the (1) handling and maintenance of mouse embryonic stem (ES) cells; (2) analysis of genomic DNA by Southern Blot; and (3) sequencing and PCR analysis of recombined genomic DNA. These experimental steps have been undertaken for the generation of new mouse models harboring cancer-associated PTEN mutations.
|Title of host publication||PTEN|
|Subtitle of host publication||Methods and Protocols|
|Editors||Leonardo Salmena, Vuk Stambolic|
|Place of Publication||New York NY USA|
|Number of pages||18|
|Publication status||Published - 2016|
|Name||Methods in Molecular Biology: Springer Protocols|