Bonnet-Garnier Amélie, Veillard Anne-Clémence, Bed'Hom Bertrand, Hayes Hélène, Britton-Davidian Janice
UMR1198 Biologie du Développement et Reproduction, INRA, F-78350, Jouy-en-Josas, France,
Methods Mol Biol. 2015;1222:83-99. doi: 10.1007/978-1-4939-1594-1_7.
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.
体细胞核移植（SCNT）成功率很低，很少超过5%。此外，体细胞核移植需要高技术技能，并且可能受到所用生物材料（卵母细胞和供体细胞质量）的影响。因此，检查供体细胞核型的正常性至关重要。至少使用G显带进行细胞遗传学分析以检测染色体数目和结构异常，从而识别染色体。在此，我们描述了进行完整细胞遗传学分析所需的经典方案，即通过G显带来识别染色体畸变，随后使用特定探针进行荧光原位杂交（FISH），以更灵敏地检测和精确识别重排。
