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.
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Acknowledgements
The authors wish to acknowledge Josette Catalan and the CeMEB Cytogenomics platform for their assistance in mouse stem cell karyotype analysis, Vincent Brochard and Sylvie Ruffini for their technical help in stem cells culturing and bovine fibroblast cultures respectively and Anne Calgaro and Alain Pinton for their advice on bovine karyotype analysis.
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Bonnet-Garnier, A., Veillard, AC., Bed’Hom, B., Hayes, H., Britton-Davidian, J. (2015). Assessing the Quality of Donor Cells: Karyotyping Methods. In: Beaujean, N., Jammes, H., Jouneau, A. (eds) Nuclear Reprogramming. Methods in Molecular Biology, vol 1222. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1594-1_7
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DOI: https://doi.org/10.1007/978-1-4939-1594-1_7
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Online ISBN: 978-1-4939-1594-1
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