[HTML][HTML] Pooling across cells to normalize single-cell RNA sequencing data with many zero counts

AT L. Lun, K Bach, JC Marioni - Genome biology, 2016 - Springer
Genome biology, 2016Springer
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 …
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.
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