RAPID GENERATION OF CHROMOSOME-SPECIFIC ALPHOID DNA PROBES USING THE POLYMERASE CHAIN-REACTION
RAPID GENERATION OF CHROMOSOME-SPECIFIC ALPHOID DNA PROBES USING THE POLYMERASE CHAIN-REACTION
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DOI：
10.1007/bf00215682
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发表时间：
1992-02-01
期刊：
影响因子：
5.3
通讯作者：
FEATHERSTONE, T
中科院分区：
文献类型：
--
作者：
DUNHAM, I;LENGAUER, C;FEATHERSTONE, T
染色体特异性α DNA探针的非同位素原位杂交已成为研究细胞周期各阶段特定人类染色体数目畸变的有力工具。在本文中，我们描述了使用聚合酶链反应（PCR）快速生成这种探针的方法，并通过荧光原位杂交证明其染色体特异性正常人中期扩散和间期核。使用针对α卫星单体保守区域的寡核苷酸引物从含有各种人类染色体的体细胞杂交细胞以及来自分选的人类染色体的DNA文库中生成染色体特异性DNA探针。α卫星单体的染色体特异性区域的寡核苷酸引物用于直接从人基因组DNA产生人染色体1、6、7、17和X的着丝粒周围异染色质的特异性DNA探针。
Non-isotopic in situ hybridization of chromosome-specific alphoid DNA probes has become a potent tool in the study of numerical aberrations of specific human chromosomes at all stages of the cell cycle. In this paper, we describe approaches for the rapid generation of such probes using the polymerase chain reaction (PCR), and demonstrate their chromosome specificity by fluorescence in situ hybridization to normal human metaphase spreads and interphase nuclei. Oligonucleotide primers for conserved regions of the alpha satellite monomer were used to generate chromosome-specific DNA probes from somatic hybrid cells containing various human chromosomes, and from DNA libraries from sorted human chromosomes. Oligonucleotide primers for chromosome-specific regions of the alpha satellite monomer were used to generate specific DNA probes for the pericentromeric heterochromatin of human chromosomes 1, 6, 7, 17 and X directly from human genomic DNA.