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非放射性原位杂交:最新技术与应用

Nonradioactive In Situ Hybridization: Recent Techniques and Applications.

作者信息

Hara Masayuki, Yamada Shozo, Hirata Kazuaki

出版信息

Endocr Pathol. 1998 Spring;9(1):21-29. doi: 10.1007/BF02739948.

Abstract

In situ hybridization (ISH) has become a standard method for the localization of nucleic acid sequences in chromosomes, single cells, and tissue sections. Nonradioactive ISH has not only eliminated the problems associated with radioactive probes but has also achieved a higher degree of resolution. Advances in probe preparation and labeling methods have facilitated the general application of ISH. In combination with immunohistochemistry, ISH can provide histological information on gene activity at the DNA, mRNA, and protein levels. Some nonradioactive ISH can simultaneously detect nucleic acid sequences in the same tissue or in a chromosome spread. Advances in ISH technology, including use of the polymerase chain reaction offer both a high sensitivity allowing detection of low levels of gene expression and the cytological localization of gene sequences.

摘要

原位杂交(ISH)已成为在染色体、单细胞和组织切片中定位核酸序列的标准方法。非放射性原位杂交不仅消除了与放射性探针相关的问题,而且还实现了更高的分辨率。探针制备和标记方法的进展促进了原位杂交的广泛应用。与免疫组织化学相结合,原位杂交可以在DNA、mRNA和蛋白质水平上提供有关基因活性的组织学信息。一些非放射性原位杂交可以同时检测同一组织或染色体铺片中的核酸序列。原位杂交技术的进展,包括聚合酶链反应的应用,既提供了高灵敏度,能够检测低水平的基因表达,又能对基因序列进行细胞学定位。

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