| Literature DB >> 17446878 |
Magdaline Costa1, Mirella Dottori, Koula Sourris, Pegah Jamshidi, Tanya Hatzistavrou, Richard Davis, Lisa Azzola, Steven Jackson, Sue Mei Lim, Martin Pera, Andrew G Elefanty, Edouard G Stanley.
Abstract
The ability to genetically modify human embryonic stem cells (HESCs) will be critical for their widespread use as a tool for understanding fundamental aspects of human biology and pathology and for their development as a platform for pharmaceutical discovery. Here, we describe a method for the genetic modification of HESCs using electroporation, the preferred method for introduction of DNA into cells in which the desired outcome is gene targeting. This report provides methods for cell amplification, electroporation, colony selection and screening. The protocol we describe has been tested on four different HESC lines, and takes approximately 4 weeks from electroporation to PCR screening of G418-resistant clones.Entities:
Mesh：
Year: 2007 PMID： 17446878 DOI： 10.1038/nprot.2007.105
Source DB: PubMed Journal: Nat Protoc ISSN： 1750-2799 Impact factor: 13.491