The dynamics of genome replication using deep sequencing
書誌事項
- 公開日
- 2013-10-01
- 権利情報
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- http://creativecommons.org/licenses/by/3.0/
- DOI
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- 10.1093/nar/gkt878
- 公開者
- Oxford University Press (OUP)
この論文をさがす
説明
Eukaryotic genomes are replicated from multiple DNA replication origins. We present complementary deep sequencing approaches to measure origin location and activity in Saccharomyces cerevisiae. Measuring the increase in DNA copy number during a synchronous S-phase allowed the precise determination of genome replication. To map origin locations, replication forks were stalled close to their initiation sites; therefore, copy number enrichment was limited to origins. Replication timing profiles were generated from asynchronous cultures using fluorescence-activated cell sorting. Applying this technique we show that the replication profiles of haploid and diploid cells are indistinguishable, indicating that both cell types use the same cohort of origins with the same activities. Finally, increasing sequencing depth allowed the direct measure of replication dynamics from an exponentially growing culture. This is the first time this approach, called marker frequency analysis, has been successfully applied to a eukaryote. These data provide a high-resolution resource and methodological framework for studying genome biology.
収録刊行物
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- Nucleic Acids Research
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Nucleic Acids Research 42 (1), e3-e3, 2013-10-01
Oxford University Press (OUP)