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[克隆DNA序列原位杂交及人类染色体鉴别染色条件的优化]

[Optimization of the conditions for the in situ hybridization of cloned DNA sequences and for the differential staining of human chromosomes].

作者信息

Iurov Iu B

出版信息

Biull Eksp Biol Med. 1984 May;97(5):595-8.

PMID:6722340
Abstract

The influence of different experimental conditions on in situ hybridization of DNA and subsequent differential staining of chromosomes was studied. The most optimal conditions for chromosomal localization of cloned repetitive DNA sequences were the lack of chromosome pretreatment with acid and RNase, reduction of the denaturation time to 30 s, carrying out of hybridization at a relatively low temperature (under 37 degrees C) at the expense of the use of formamide, addition to the hybridization mixture of 10% of dextran sulfate-500. The conditions indicated permit obtaining on radioautographs the G- and C-segmentation of human chromosomes.

摘要

研究了不同实验条件对DNA原位杂交及随后染色体差异染色的影响。克隆重复DNA序列染色体定位的最适宜条件为:不进行酸和核糖核酸酶的染色体预处理;将变性时间缩短至30秒;以使用甲酰胺为代价,在相对低温(37摄氏度以下)下进行杂交;在杂交混合物中添加10%的硫酸葡聚糖-500。所述条件可在放射自显影片上获得人类染色体的G带和C带。

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