首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines.
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Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines.

机译:与Alu和L1聚合酶链反应探针的荧光原位杂交技术可快速鉴定杂交细胞系中人类染色体的特征。

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摘要

Human-rodent hybrid cell lines have been analyzed with regard to their human DNA content by using various DNA probe sets, derived from the hybrids, for in situ hybridization to normal human metaphase chromosome spreads. Total genomic hybrid DNA was compared with probe sets of hybrid DNA that were highly enriched in human sequences. The latter probes were obtained by amplification through the polymerase chain reaction (PCR) using oligonucleotide primers directed to human specific subsequences of the interspersed repetitive sequences Alu and L1. Previously unidentified chromosomal material within hybrid lines was characterized with speed and precision. It is demonstrated that the complete human complement of hybrid lines can be rapidly assessed by comparing the data obtained with the Alu-PCR products with the results from the L1-PCR products or from the genomic hybrid DNA. This approach using interspersed repetitive sequence-PCR products is simple and fast and also provides an alternative way of generating complex DNA probe sets for the specific delineation of entire chromosomes or subchromosomal regions by in situ hybridization.
机译:已经通过使用衍生自杂种的各种DNA探针组对人-啮齿动物杂交细胞系的人类DNA含量进行了分析,以便与正常人类中期染色体的传播进行原位杂交。将总基因组杂合DNA与高度富集于人类序列的杂合DNA探针组进行比较。后者的探针是通过使用针对散布的重复序列Alu和L1的人特异性亚序列的寡核苷酸引物通过聚合酶链反应(PCR)扩增而获得的。杂交品系中以前未鉴定的染色体材料具有快速和精确的特征。通过将Alu-PCR产物获得的数据与L1-PCR产物或基因组杂交DNA的结果进行比较,可以快速评估杂种系的完整人类补体。这种使用散布的重复序列-PCR产物的方法既简单又快速,并且还提供了另一种方法,可通过原位杂交生成复杂的DNA探针组,以特定地描绘整个染色体或亚染色体区域。

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