Статья

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

J. Gregory CaporasoDepartment of Computer Science, Northern Arizona University , Flagstaff, AZ , USAChristian L. LauberCooperative Institute for Research in Environmental Sciences, UCB 216, University of Colorado , Boulder, CO , USAWilliam A. WaltersDepartment of Molecular, Cellular and Developmental Biology, UCB 347, University of Colorado , Boulder, CO , USADonna Berg-LyonsCooperative Institute for Research in Environmental Sciences, UCB 216, University of Colorado , Boulder, CO , USAJames S. HuntleyColorado Initiative in Molecular Biotechnology, UCB 347, University of Colorado , Boulder, CO , USANoah FiererCooperative Institute for Research in Environmental Sciences, UCB 216, University of Colorado , Boulder, CO , USASarah M. OwensArgonne National Laboratory , Argonne, IL , USAJason BetleyIllumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UKLouise FraserIllumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UKMarkus BauerIllumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UKNiall GormleyIllumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UKJack A. GilbertArgonne National Laboratory , Argonne, IL , USAGeoff SmithIllumina Cambridge Ltd., Chesterford Research Park, Saffron Walden , Essex , UKRob KnightDepartment of Chemistry and Biochemistry, UCB 215, University of Colorado , Boulder, CO , USA

2012en

ABI

Аннотация

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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Идентификаторы

DOI: 10.1038/ismej.2012.8

Цитирования и источники

Цитирований: 7Использованных источников: 0

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