Quantitative single-cell RNA-seq with unique molecular identifiers
Nature methods, 2014•nature.com
Single-cell RNA sequencing (RNA-seq) is a powerful tool to reveal cellular heterogeneity,
discover new cell types and characterize tumor microevolution. However, losses in cDNA
synthesis and bias in cDNA amplification lead to severe quantitative errors. We show that
molecular labels—random sequences that label individual molecules—can nearly eliminate
amplification noise, and that microfluidic sample preparation and optimized reagents
produce a fivefold improvement in mRNA capture efficiency.
discover new cell types and characterize tumor microevolution. However, losses in cDNA
synthesis and bias in cDNA amplification lead to severe quantitative errors. We show that
molecular labels—random sequences that label individual molecules—can nearly eliminate
amplification noise, and that microfluidic sample preparation and optimized reagents
produce a fivefold improvement in mRNA capture efficiency.
Abstract
Single-cell RNA sequencing (RNA-seq) is a powerful tool to reveal cellular heterogeneity, discover new cell types and characterize tumor microevolution. However, losses in cDNA synthesis and bias in cDNA amplification lead to severe quantitative errors. We show that molecular labels—random sequences that label individual molecules—can nearly eliminate amplification noise, and that microfluidic sample preparation and optimized reagents produce a fivefold improvement in mRNA capture efficiency.
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