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引用次数: 27
摘要
引物原位标记(Primed in situ labeling, PRINS)是一种灵敏、特异的技术,可用于定位传统FISH检测不到的单拷贝基因和DNA片段。利用PRINS,我们将SRY基因定位到Yp11.31p11.32, SOX3基因定位到Xq26q27。基因座特异性寡核苷酸引物原位退火,延伸到固定在显微镜载玻片上的染色体制剂上,存在有dATP、dCTP、dGTP、dTTP、生物素-16- dutp、Tris-HCl、KCl、MgCl2、BSA和Taq DNA聚合酶。在中期扩散和间期细胞核中检测到荧光信号。我们的方法对于一般的单拷贝基因来说是有价值的。
Chromosomal localization of single copy genes SRY and SOX3 by primed in situ labeling (PRINS).
Primed in situ labeling (PRINS) is a sensitive and specific technique that can be used for the localization of single copy genes and DNA segments that are too small to be detected by conventional FISH. With PRINS, we physically localized the SRY gene to Yp11.31p11.32 and the SOX3 gene to Xq26q27. Locus-specific oligonucleotide primers were annealed in situ and extended on chromosome preparations fixed on microscope slides, in the presence of dATP, dCTP, dGTP, dTTP, biotin-16-dUTP, Tris-HCl, KCl, MgCl2, BSA, and Taq DNA polymerase. Fluorescent signals were detected in metaphase spreads and interphase nuclei. Our method may prove valuable for use with single copy genes in general.