title: Assessing the quality of donor cells: karyotyping methods.
abstract: Somatic cell nuclear transfer (SCNT) has a low success rate that rarely exceeds 5 %. Moreover, SCNT requires highly technical skills and may be influenced by the biological material used (oocyte and donor cell quality). Hence, it is crucial to check the normality of the donor cell's karyotype. Numerical and structural chromosome abnormalities are detected by cytogenetic analysis at minimum using G-banding to identify the chromosomes. Here, we describe the classical protocols that are needed to perform complete cytogenetic analyses, i.e., G-banding to identify chromosome aberrations, followed by Fluorescent In Situ Hybridization (FISH) of specific probes for a more sensitive detection and precise identification of the rearrangement.
authors.affiliation: UMR1198 Biologie du Développement et Reproduction, INRA, F-78350, Jouy-en-Josas, France, amelie.bonnet-garnier@jouy.inra.fr.
authors.forename: Amélie
authors.initials: A
authors.lastname: Bonnet-Garnier
authors.forename: Anne-Clémence
authors.initials: AC
authors.lastname: Veillard
authors.forename: Bertrand
authors.initials: B
authors.lastname: Bed'Hom
authors.forename: Hélène
authors.initials: H
authors.lastname: Hayes
authors.forename: Janice
authors.initials: J
authors.lastname: Britton-Davidian
mesh_terms: D000818:Animals; D002417:Cattle; D002869:Chromosome Aberrations; D002871:Chromosome Banding / Q000379:methods*; D053595:Embryonic Stem Cells; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007621:Karyotyping / Q000379:methods; D008677:Metaphase; D051379:Mice; D014019:Tissue Donors; D014357:Trypsin / Q000737:chemistry
chemical_list: D014357:Trypsin