Normalizing single-cell RNA sequencing data: challenges and opportunities

CA Vallejos, D Risso, A Scialdone, S Dudoit… - Nature …, 2017 - nature.com
Nature methods, 2017nature.com
Single-cell transcriptomics is becoming an important component of the molecular biologist's
toolkit. A critical step when analyzing data generated using this technology is normalization.
However, normalization is typically performed using methods developed for bulk RNA
sequencing or even microarray data, and the suitability of these methods for single-cell
transcriptomics has not been assessed. We here discuss commonly used normalization
approaches and illustrate how these can produce misleading results. Finally, we present …
Abstract
Single-cell transcriptomics is becoming an important component of the molecular biologist's toolkit. A critical step when analyzing data generated using this technology is normalization. However, normalization is typically performed using methods developed for bulk RNA sequencing or even microarray data, and the suitability of these methods for single-cell transcriptomics has not been assessed. We here discuss commonly used normalization approaches and illustrate how these can produce misleading results. Finally, we present alternative approaches and provide recommendations for single-cell RNA sequencing users.
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