Sister chromatid differentiation with biotin-dUTP

被引：12

作者：

Bruckmann, E

Wojcik, A

Obe, G ^{[1
]}

机构：

[1] Univ Essen Gesamthsch, Inst Genet, D-45117 Essen, Germany

[2] Inst Nucl Chem & Technol, Dept Radiol & Hlth Protect, PL-00981 Warsaw, Poland

来源：

CHROMOSOME RESEARCH | 1999年 / 7卷 / 03期

关键词：

biotin-dUTP; CHO; fluorescence microscopy; SCE; sister chromatid exchanges;

D O I：

10.1023/A:1009295131877

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The method of choice to differentiate sister chromatids is to incorporate BrdU in replicating DNA. The disadvantage of BrdU is that its spontaneous or induced radicalization may itself lead to sister chromatid exchanges. Biotin-labelled dUTP is a widely used thymidine analogon for labelling isolated DNA. Its chemical structure suggests that, in contrast to BrdU, it does not give rise to radical formation. We electroporated proliferating Chinese hamster ovary (CHO) cells in the presence of biotin-dUTP which was subsequently detected in metaphase cells with TRITC-conjugated avidin. Microscopic analysis of second mitoses after labelling revealed a clear differential staining of sister chromatids. Thus substitution of thymidine with biotin-dUTP is another method to analyse SCE.

引用

页码：185 / 189

页数：5

共 14 条

[1] IMMUNOFLUORESCENT VISUALIZATION OF DNA-REPLICATION SITES WITHIN NUCLEI OF CHINESE-HAMSTER OVARY CELLS [J].