Pellestor Franck, Andréo Brigitte, Puechberty Jacques, Lefort Geneviève, Sarda Pierre
Institute of Human Genetics, CNRS UPR 1142, Montpellier, France.
Methods Mol Biol. 2006;334:49-59. doi: 10.1385/1-59745-068-5:49.
Based on the direct in situ mixing of the colors of different fluorochromes (fluorescein isothiocyanate, tetramethylrhodamine isothiocyanate, Cascade Blue) incorporated in sequential primed in situ labeling (PRINS) reactions, a new multicolor PRINS procedure is described, allowing the rapid and distinct in situ labeling of three or four human chromosomes. Each PRINS reaction consists of a unique 5-min step for annealing and elongation. In combination with the 0.5 M NaOH pretreatment for simultaneous in situ denaturation and decondensation of sperm nuclei, this technique has been adapted to human sperm nuclei for the direct assessment of aneuploidy.
基于在连续引物原位标记（PRINS）反应中掺入的不同荧光染料（异硫氰酸荧光素、异硫氰酸四甲基罗丹明、级联蓝）颜色的直接原位混合，描述了一种新的多色PRINS程序，可对三或四条人类染色体进行快速且清晰的原位标记。每个PRINS反应包括一个独特的5分钟退火和延伸步骤。结合用于精子核原位变性和去浓缩的0.5M NaOH预处理，该技术已应用于人类精子核以直接评估非整倍性。
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