Barcoding bias in high-throughput multiplex sequencing of miRNA

被引：88

作者：

Alon, Shahar ^{[2
]}

Vigneault, Francois ^{[3
,4
,5
]}

Eminaga, Seda ^{[3
]}

Christodoulou, Danos C. ^{[3
]}

Seidman, Jonathan G. ^{[3
]}

Church, George M. ^{[3
,4
]}

Eisenberg, Eli ^{[1
]}

机构：

[1] Tel Aviv Univ, Raymond & Beverly Sackler Sch Phys & Astron, IL-69978 Tel Aviv, Israel

[2] Tel Aviv Univ, George S Wise Fac Life Sci, Dept Neurobiol, IL-69978 Tel Aviv, Israel

[3] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02115 USA

[4] Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA

[5] Ragon Inst MGH MIT & Harvard, Boston, MA 02129 USA

来源：

GENOME RESEARCH | 2011年 / 21卷 / 09期

基金：

加拿大健康研究院;

关键词：

MICRORNAS; IDENTIFICATION; GENOMICS;

D O I：

10.1101/gr.121715.111

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Second-generation sequencing is gradually becoming the method of choice for miRNA detection and expression profiling. Given the relatively small number of miRNAs and improvements in DNA sequencing technology, studying miRNA expression profiles of multiple samples in a single flow cell lane becomes feasible. Multiplexing strategies require marking each miRNA library with a DNA barcode. Here we report that barcodes introduced through adapter ligation confer significant bias on miRNA expression profiles. This bias is much higher than the expected Poisson noise and masks significant expression differences between miRNA libraries. This bias can be eliminated by adding barcodes during PCR amplification of libraries. The accuracy of miRNA expression measurement in multiplexed experiments becomes a function of sample number.

引用

页码：1506 / 1511

页数：6

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