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SubSeq: determining appropriate sequencing depth through efficient read subsampling.


ABSTRACT:

Motivation

Next-generation sequencing experiments, such as RNA-Seq, play an increasingly important role in biological research. One complication is that the power and accuracy of such experiments depend substantially on the number of reads sequenced, so it is important and challenging to determine the optimal read depth for an experiment or to verify whether one has adequate depth in an existing experiment.

Results

By randomly sampling lower depths from a sequencing experiment and determining where the saturation of power and accuracy occurs, one can determine what the most useful depth should be for future experiments, and furthermore, confirm whether an existing experiment had sufficient depth to justify its conclusions. We introduce the subSeq R package, which uses a novel efficient approach to perform this subsampling and to calculate informative metrics at each depth.

Availability and implementation

The subSeq R package is available at http://github.com/StoreyLab/subSeq/.

SUBMITTER: Robinson DG 

PROVIDER: S-EPMC4296149 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature

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subSeq: determining appropriate sequencing depth through efficient read subsampling.

Robinson David G DG   Storey John D JD  

Bioinformatics (Oxford, England) 20140903 23


<h4>Motivation</h4>Next-generation sequencing experiments, such as RNA-Seq, play an increasingly important role in biological research. One complication is that the power and accuracy of such experiments depend substantially on the number of reads sequenced, so it is important and challenging to determine the optimal read depth for an experiment or to verify whether one has adequate depth in an existing experiment.<h4>Results</h4>By randomly sampling lower depths from a sequencing experiment and  ...[more]

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