期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 425, 期 23, 页码 4845-4855出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2013.03.040
关键词
DNA combing; replication fork speed; inter-origin distance; inter-termination distance; fork asymmetry
资金
- Institut National du Cancer [2009-1-PLBIO-10-IC-1]
- Agence Nationale de la Recherche [ANR-09-GENO-000/repinsCFS]
- Association pour la Recherche sur le Cancer [SL220100601348, 8514]
- MENRT graduate student fellowship
- Association pour la Recherche sur le Cancer
- Institut National du Cancer
The factors that govern replication programs are still poorly identified in metazoans, especially in mammalian cells. Thanks to molecular combing, the dynamics of DNA replication can be assessed at the genome-scale level from the cumulative analysis of single DNA fibers. This technique notably enables measurement of replication fork speed and fork asymmetry and that of distances separating either initiation or termination events. The results presented here aim to evaluate requirements critical to accurate measurement of replication parameters by molecular combing. We show that sample size, fiber length and DNA counterstaining are crucial to gain robust information concerning replication dynamics. Our results thus provide a methodological frame to investigate the DNA replication program through molecular combing analyses. (C) 2013 Elsevier Ltd. All rights reserved.
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