BackgroundEpigenomic profiling assays such as ChIP-seq have been widely used to map the genome-wide enrichment profiles of chromatin-associated proteins and posttranslational histone modifications. Sequencing depth is a key parameter in experimental design and quality control. However, due to variable sequencing depth requirements across experimental conditions, it can be challenging to determine optimal sequencing depth, particularly for projects involving multiple targets or cell types.ResultsWe developed the peaksat R package to provide target read depth estimates for epigenomic experiments based on the analysis of peak saturation curves. We applied peaksat to establish the distinctive read depth requirements for ChIP-seq studies of histone modifications in different cell lines. Using peaksat, we were able to estimate the target read depth required per library to obtain high-quality peak calls for downstream analysis. In addition, peaksat was applied to other sequence-enrichment methods including CUT&RUN and ATAC-seq.Conclusionpeaksat addresses a need for researchers to make informed decisions about whether their sequencing data has been generated to an adequate depth and subsequently sufficient meaningful peaks, and failing that, how many more reads would be required per library. peaksat is applicable to other sequence-based methods that include calling peaks in their analysis.
机构:
Walter & Eliza Hall Inst Med Res, 1G Royal Parade, Parkville, Vic 3052, Australia
Univ Melbourne, Dept Med Biol, Parkville, Vic 3010, AustraliaWalter & Eliza Hall Inst Med Res, 1G Royal Parade, Parkville, Vic 3052, Australia
Lun, Aaron T. L.
Smyth, Gordon K.
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Walter & Eliza Hall Inst Med Res, 1G Royal Parade, Parkville, Vic 3052, Australia
Univ Melbourne, Dept Math & Stat, Parkville, Vic 3010, AustraliaWalter & Eliza Hall Inst Med Res, 1G Royal Parade, Parkville, Vic 3052, Australia
机构:
Univ N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Univ Wisconsin, Dept Stat, Madison, WI 53706 USA
Univ Wisconsin, Dept Biostat & Med Informat, Madison, WI 53706 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Kuan, Pei Fen
Chung, Dongjun
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Univ Wisconsin, Dept Stat, Madison, WI 53706 USA
Univ Wisconsin, Dept Biostat & Med Informat, Madison, WI 53706 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Chung, Dongjun
Pan, Guangjin
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Chinese Acad Sci, Guangzhou Inst Biomed, Guangzhou 510530, Guangdong, Peoples R China
Chinese Acad Sci, Guangzhou Inst Hlth, Guangzhou 510530, Guangdong, Peoples R China
Univ Wisconsin, Morgridge Inst Res, Madison, WI 53715 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Pan, Guangjin
Thomson, James A.
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Univ Wisconsin, Dept Anat, Genome Ctr Wisconsin, Madison, WI 53715 USA
Univ Wisconsin, Morgridge Inst Res, Madison, WI 53715 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Thomson, James A.
Stewart, Ron
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Univ Wisconsin, Morgridge Inst Res, Madison, WI 53715 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA
Stewart, Ron
Keles, Suenduez
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Univ Wisconsin, Dept Stat, Madison, WI 53706 USA
Univ Wisconsin, Dept Biostat & Med Informat, Madison, WI 53706 USAUniv N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA