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Sensitivity to sequencing depth in single-cell cancer genomics.


ABSTRACT: Querying cancer genomes at single-cell resolution is expected to provide a powerful framework to understand in detail the dynamics of cancer evolution. However, given the high costs currently associated with single-cell sequencing, together with the inevitable technical noise arising from single-cell genome amplification, cost-effective strategies that maximize the quality of single-cell data are critically needed. Taking advantage of previously published single-cell whole-genome and whole-exome cancer datasets, we studied the impact of sequencing depth and sampling effort towards single-cell variant detection.Five single-cell whole-genome and whole-exome cancer datasets were independently downscaled to 25, 10, 5, and 1× sequencing depth. For each depth level, ten technical replicates were generated, resulting in a total of 6280 single-cell BAM files. The sensitivity of variant detection, including structural and driver mutations, genotyping, clonal inference, and phylogenetic reconstruction to sequencing depth was evaluated using recent tools specifically designed for single-cell data.Altogether, our results suggest that for relatively large sample sizes (25 or more cells) sequencing single tumor cells at depths >?5× does not drastically improve somatic variant discovery, characterization of clonal genotypes, or estimation of single-cell phylogenies.We suggest that sequencing multiple individual tumor cells at a modest depth represents an effective alternative to explore the mutational landscape and clonal evolutionary patterns of cancer genomes.

SUBMITTER: Alves JM 

PROVIDER: S-EPMC5901877 | biostudies-other | 2018 Apr

REPOSITORIES: biostudies-other

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Sensitivity to sequencing depth in single-cell cancer genomics.

Alves João M JM   Posada David D  

Genome medicine 20180416 1


<h4>Background</h4>Querying cancer genomes at single-cell resolution is expected to provide a powerful framework to understand in detail the dynamics of cancer evolution. However, given the high costs currently associated with single-cell sequencing, together with the inevitable technical noise arising from single-cell genome amplification, cost-effective strategies that maximize the quality of single-cell data are critically needed. Taking advantage of previously published single-cell whole-gen  ...[more]

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