Molecular Combing of Single DNA Molecules on the 10 Megabase Scale

被引：29

作者：

Kaykov, Atanas ^{[1
]}

Taillefumier, Thibaud ^{[2
]}

Bensimon, Aaron ^{[3
]}

Nurse, Paul ^{[1
,4
]}

机构：

[1] Rockefeller Univ, New York, NY 10065 USA

[2] Princeton Univ, Lewis Sigler Inst Integrat Genom, Princeton, NJ 08544 USA

[3] Genom Vis, F-92220 Bagneux, France

[4] Francis Crick Inst, Lincolns Inn Fields Labs, London WC2A 3LY, England

来源：

SCIENTIFIC REPORTS | 2016年 / 6卷

基金：

英国惠康基金; 英国医学研究理事会;

关键词：

REPLICATION ORIGINS; GENOME-WIDE; SCHIZOSACCHAROMYCES-POMBE; FORK PROGRESSION; YEAST; REVEALS; DYNAMICS; CELLS; IDENTIFICATION; TRANSCRIPTION;

D O I：

10.1038/srep19636

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

DNA combing allows the investigation of DNA replication on genomic single DNA molecules, but the lengths that can be analysed have been restricted to molecules of 200-500 kb. We have improved the DNA combing procedure so that DNA molecules can be analysed up to the length of entire chromosomes in fission yeast and up to 12 Mb fragments in human cells. Combing multi-Mb-scale DNA molecules revealed previously undetected origin clusters in fission yeast and shows that in human cells replication origins fire stochastically forming clusters of fired origins with an average size of 370 kb. We estimate that a single human cell forms around 3200 clusters at mid S-phase and fires approximately 100,000 origins to complete genome duplication. The procedure presented here will be adaptable to other organisms and experimental conditions.

引用

页数：9

共 42 条

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