An improved dual-indexing approach for multiplexed 16S rRNA gene sequencing on the Illumina MiSeq platform View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2014-02-24

AUTHORS

Douglas W Fadrosh, Bing Ma, Pawel Gajer, Naomi Sengamalay, Sandra Ott, Rebecca M Brotman, Jacques Ravel

ABSTRACT

BackgroundTo take advantage of affordable high-throughput next-generation sequencing technologies to characterize microbial community composition often requires the development of improved methods to overcome technical limitations inherent to the sequencing platforms. Sequencing low sequence diversity libraries such as 16S rRNA amplicons has been problematic on the Illumina MiSeq platform and often generates sequences of suboptimal quality.ResultsHere we present an improved dual-indexing amplification and sequencing approach to assess the composition of microbial communities from clinical samples using the V3-V4 region of the 16S rRNA gene on the Illumina MiSeq platform. We introduced a 0 to 7 bp “heterogeneity spacer” to the index sequence that allows an equal proportion of samples to be sequenced out of phase.ConclusionsOur approach yields high quality sequence data from 16S rRNA gene amplicons using both 250 bp and 300 bp paired-end MiSeq protocols and provides a flexible and cost-effective sequencing option. More... »

PAGES

6

References to SciGraph publications

Journal

TITLE

Microbiome

ISSUE

1

VOLUME

2

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/2049-2618-2-6

DOI

http://dx.doi.org/10.1186/2049-2618-2-6

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1017337669

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24558975


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186 Institute for Genome Sciences, Department of Microbiology and Immunology, University of Maryland School of Medicine, W. Baltimore Street, 801, 21201, Baltimore, MD, USA
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