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- Publisher Website: 10.1093/nar/gku178
- Scopus: eid_2-s2.0-84901303566
- PMID: 24598259
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Article: Impact of sequencing depth in ChIP-seq experiments
Title | Impact of sequencing depth in ChIP-seq experiments |
---|---|
Authors | |
Issue Date | 2014 |
Citation | Nucleic Acids Research, 2014, v. 42, n. 9 How to Cite? |
Abstract | In a chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) experiment, an important consideration in experimental design is the minimum number of sequenced reads required to obtain statistically significant results. We present an extensive evaluation of the impact of sequencing depth on identification of enriched regions for key histone modifications (H3K4me3, H3K36me3, H3K27me3 and H3K9me2/me3) using deep-sequenced datasets in human and fly. We propose to define sufficient sequencing depth as the number of reads at which detected enrichment regions increase <1% for an additional million reads. Although the required depth depends on the nature of the mark and the state of the cell in each experiment, we observe that sufficient depth is often reached at <20 million reads for fly. For human, there are no clear saturation points for the examined datasets, but our analysis suggests 40-50 million reads as a practical minimum for most marks. We also devise a mathematical model to estimate the sufficient depth and total genomic coverage of a mark. Lastly, we find that the five algorithms tested do not agree well for broad enrichment profiles, especially at lower depths. Our findings suggest that sufficient sequencing depth and an appropriate peak-calling algorithm are essential for ensuring robustness of conclusions derived from ChIP-seq data. © 2014 The Author(s) 2014. |
Persistent Identifier | http://hdl.handle.net/10722/262652 |
ISSN | 2023 Impact Factor: 16.6 2023 SCImago Journal Rankings: 7.048 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Jung, Youngsook L. | - |
dc.contributor.author | Luquette, Lovelace J. | - |
dc.contributor.author | Ho, Joshua W K | - |
dc.contributor.author | Ferrari, Francesco | - |
dc.contributor.author | Tolstorukov, Michael | - |
dc.contributor.author | Minoda, Aki | - |
dc.contributor.author | Issner, Robbyn | - |
dc.contributor.author | Epstein, Charles B. | - |
dc.contributor.author | Karpen, Gary H. | - |
dc.contributor.author | Kuroda, Mitzi I. | - |
dc.contributor.author | Park, Peter J. | - |
dc.date.accessioned | 2018-10-08T02:46:38Z | - |
dc.date.available | 2018-10-08T02:46:38Z | - |
dc.date.issued | 2014 | - |
dc.identifier.citation | Nucleic Acids Research, 2014, v. 42, n. 9 | - |
dc.identifier.issn | 0305-1048 | - |
dc.identifier.uri | http://hdl.handle.net/10722/262652 | - |
dc.description.abstract | In a chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) experiment, an important consideration in experimental design is the minimum number of sequenced reads required to obtain statistically significant results. We present an extensive evaluation of the impact of sequencing depth on identification of enriched regions for key histone modifications (H3K4me3, H3K36me3, H3K27me3 and H3K9me2/me3) using deep-sequenced datasets in human and fly. We propose to define sufficient sequencing depth as the number of reads at which detected enrichment regions increase <1% for an additional million reads. Although the required depth depends on the nature of the mark and the state of the cell in each experiment, we observe that sufficient depth is often reached at <20 million reads for fly. For human, there are no clear saturation points for the examined datasets, but our analysis suggests 40-50 million reads as a practical minimum for most marks. We also devise a mathematical model to estimate the sufficient depth and total genomic coverage of a mark. Lastly, we find that the five algorithms tested do not agree well for broad enrichment profiles, especially at lower depths. Our findings suggest that sufficient sequencing depth and an appropriate peak-calling algorithm are essential for ensuring robustness of conclusions derived from ChIP-seq data. © 2014 The Author(s) 2014. | - |
dc.language | eng | - |
dc.relation.ispartof | Nucleic Acids Research | - |
dc.rights | This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. | - |
dc.title | Impact of sequencing depth in ChIP-seq experiments | - |
dc.type | Article | - |
dc.description.nature | published_or_final_version | - |
dc.identifier.doi | 10.1093/nar/gku178 | - |
dc.identifier.pmid | 24598259 | - |
dc.identifier.scopus | eid_2-s2.0-84901303566 | - |
dc.identifier.volume | 42 | - |
dc.identifier.issue | 9 | - |
dc.identifier.spage | null | - |
dc.identifier.epage | null | - |
dc.identifier.eissn | 1362-4962 | - |
dc.identifier.isi | WOS:000336495400001 | - |
dc.identifier.issnl | 0305-1048 | - |