Characterization of Targeted Mouse Embryonic Stem Cell Chromosomes : Karyotyping and Fluorescence In Situ Hybridization of Metaphase Spreads

Diana Hernandez; Ian R Phillips; Elizabeth A Shephard

doi:10.1385/1-59259-998-2:321

Characterization of Targeted Mouse Embryonic Stem Cell Chromosomes : Karyotyping and Fluorescence In Situ Hybridization of Metaphase Spreads

Methods Mol Biol. 2006:320:321-327. doi: 10.1385/1-59259-998-2:321.

Authors

Diana Hernandez¹, Ian R Phillips¹, Elizabeth A Shephard²

Affiliations

¹ Department of Biochemistry and Molecular Biology, University College London, London, UK.
² School of Biological Sciences, Queen Mary, University of London, London, UK.

PMID: 27699675
DOI: 10.1385/1-59259-998-2:321

Abstract

The manipulation of genes in mouse embryonic stem (ES) cells can result in chromosome abnormalities. This chapter describes methods for karyotyping of the manipulated ES cell line before injection into blastocysts and the use of fluorescence in situ hybridization to confirm the deletion of a targeted gene. The method is illustrated by describing how an ES cell line targeted for the deletion of Fmo genes was characterized.

Keywords: Flavin-containing monooxygenase genes; Fmo1; Fmo2; Fmo4; chromosome 1; embryonic stem cells; gene deletion; karyotyping.