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. 2014 Feb;11(2):163-6.
doi: 10.1038/nmeth.2772. Epub 2013 Dec 22.

Quantitative single-cell RNA-seq with unique molecular identifiers

Saiful Islam  1 , Amit Zeisel  1 , Simon Joost  2 , Gioele La Manno  1 , Pawel Zajac  1 , Maria Kasper  2 , Peter Lönnerberg  1 , Sten Linnarsson  1
Affiliations

Quantitative single-cell RNA-seq with unique molecular identifiers

Saiful Islam et al. Nat Methods. 2014 Feb.

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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References

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