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带有独特分子标识符的定量单细胞 RNA-seq。

Quantitative single-cell RNA-seq with unique molecular identifiers.

机构信息

Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.

Department of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden.

出版信息

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

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.

摘要

单细胞 RNA 测序(RNA-seq)是揭示细胞异质性、发现新细胞类型和描绘肿瘤微进化的有力工具。然而,cDNA 合成的损失和 cDNA 扩增的偏倚导致严重的定量误差。我们表明,分子标签(标记单个分子的随机序列)几乎可以消除扩增噪声,并且微流控样品制备和优化试剂可将 mRNA 捕获效率提高五倍。

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