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温度对原位杂交检测特定DNA序列及其染色体分布的影响。

Influence of temperature on the detectibility and chromosomal distribution of specific DNA sequences by in situ hybridisation.

作者信息

Moar M H, Purdom I F, Jones K W

出版信息

Chromosoma. 1975 Dec 29;53(4):345-59. doi: 10.1007/BF00294082.

Abstract

Hybridising certain AT-rich satellite complementary RNAs (cRNAs) to their homologous chromosomal DNA sequences at different temperatures of incubation results in a different dispersion of autoradiographic label throughout the karyotypes. The temperature at which most label, or cRNA-DNA hybrid formation, exists corresponds to the optimal rate temperature for the hybridisation of these same satellite cRNA-DNA hybrids as determined by RNA excess filter hybridisation. It is likely that the in situ hybridisation results can therefore be explained by the fact that there is a similar temperature-dependence on the rate of hybrid formation for both in situ and RNA excess hybridisation. This should have important implications for the designing of in situ hybridisation experiments in general.

摘要

在不同的孵育温度下,将某些富含AT的卫星互补RNA(cRNA)与其同源染色体DNA序列杂交,会导致放射自显影标记在整个核型中的分布不同。存在大多数标记或cRNA-DNA杂交形成的温度,与通过RNA过量滤膜杂交确定的这些相同卫星cRNA-DNA杂交体杂交的最佳速率温度相对应。因此,原位杂交结果很可能可以用以下事实来解释:原位杂交和RNA过量杂交在杂交形成速率上都存在类似的温度依赖性。总体而言,这对原位杂交实验的设计应该具有重要意义。

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