NDLI: Barcoding bias in high-throughput multiplex sequencing of miRNA.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Alon, Shahar Vigneault, Francois Eminaga, Seda Christodoulou, Danos C. Seidman, Jonathan G. Church, George M. Eisenberg, Eli
Description	Country affiliation: Israel Author Affiliation: Alon S ( Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv, Israel.)
Abstract	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.
File Format	HTM / HTML
ISSN	10889051
e-ISSN	15495469
DOI	10.1101/gr.121715.111
Journal	Genome Research
Issue Number	9
Volume Number	21
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2011-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Genetics Discipline Genomics High-throughput Nucleotide Sequencing Micrornas Metabolism Sequence Analysis, Rna Animals Bias (epidemiology) Cluster Analysis Gene Expression Profiling Mice Sequence Tagged Sites Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s.
Content Type	Text
Resource Type	Article
Subject	Genetics Genetics (clinical)

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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