Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms

ISME Journal - Tập 6 Số 8 - Trang 1621-1624 - 2012
J. Gregory Caporaso1, Christian L. Lauber2, William A. Walters3, Donna Berg-Lyons2, James S Huntley4, Noah Fierer2,5, Sarah M. Owens6, Jason Betley7, Louise Fraser7, Markus Bauer7, Niall Gormley7, Jack A. Gilbert8,9, Geoff Smith7, Rob Knight10,11
1Department of Computer Science, Northern Arizona University , Flagstaff, AZ , USA
2Cooperative Institute for Research in Environmental Sciences, UCB 216, University of Colorado , Boulder, CO , USA
3Department of Molecular, Cellular and Developmental Biology, UCB 347, University of Colorado , Boulder, CO , USA
4Colorado Initiative in Molecular Biotechnology, UCB 347, University of Colorado , Boulder, CO , USA
5Department of Ecology and Evolutionary Biology, UCB 334, University of Colorado , Boulder, Colorado , USA
6Argonne National Laboratory, Argonne, IL, USA
7Illumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UK
8Department of Ecology and Evolution, University of Chicago , Chicago, IL , USA
9Department of Ecology and Evolution, University of Chicago, Chicago, IL, USA
10Department of Chemistry and Biochemistry, UCB 215, University of Colorado , Boulder, CO , USA
11Howard Hughes Medical Institute, University of Colorado at Boulder, UCB 215 , Boulder, CO , USA

Tóm tắt

Abstract

DNA sequencing continues to decrease in cost with the Illumina HiSeq2000 generating up to 600 Gb of paired-end 100 base reads in a ten-day run. Here we present a protocol for community amplicon sequencing on the HiSeq2000 and MiSeq Illumina platforms, and apply that protocol to sequence 24 microbial communities from host-associated and free-living environments. A critical question as more sequencing platforms become available is whether biological conclusions derived on one platform are consistent with what would be derived on a different platform. We show that the protocol developed for these instruments successfully recaptures known biological results, and additionally that biological conclusions are consistent across sequencing platforms (the HiSeq2000 versus the MiSeq) and across the sequenced regions of amplicons.

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Tài liệu tham khảo

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ISME Journal

Tập 6 Số 8

1621-1624

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