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Pooling across cells to normalize single-cell RNA sequencing data with many zero counts.


ABSTRACT: Normalization of single-cell RNA sequencing data is necessary to eliminate cell-specific biases prior to downstream analyses. However, this is not straightforward for noisy single-cell data where many counts are zero. We present a novel approach where expression values are summed across pools of cells, and the summed values are used for normalization. Pool-based size factors are then deconvolved to yield cell-based factors. Our deconvolution approach outperforms existing methods for accurate normalization of cell-specific biases in simulated data. Similar behavior is observed in real data, where deconvolution improves the relevance of results of downstream analyses.

SUBMITTER: Lun AT 

PROVIDER: S-EPMC4848819 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature

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Pooling across cells to normalize single-cell RNA sequencing data with many zero counts.

Lun Aaron T L AT   Bach Karsten K   Marioni John C JC  

Genome biology 20160427


Normalization of single-cell RNA sequencing data is necessary to eliminate cell-specific biases prior to downstream analyses. However, this is not straightforward for noisy single-cell data where many counts are zero. We present a novel approach where expression values are summed across pools of cells, and the summed values are used for normalization. Pool-based size factors are then deconvolved to yield cell-based factors. Our deconvolution approach outperforms existing methods for accurate nor  ...[more]

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