Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex

We constructed error-correcting DNA barcodes that allow one run of a massively parallel pyrosequencer to process up to 1,544 samples simultaneously. Using these barcodes we processed bacterial 16S rRNA gene sequences representing microbial communities in 286 environmental samples, corrected 92% of sample assignment errors, and thus characterized nearly as many 16S rRNA genes as have been sequenced to date by Sanger sequencing.

[1]  N. Pace A molecular view of microbial diversity and the biosphere. , 1997, Science.

[2]  Philip Hugenholtz,et al.  Microbial Diversity in a Hydrocarbon- and Chlorinated-Solvent-Contaminated Aquifer Undergoing Intrinsic Bioremediation , 1998, Applied and Environmental Microbiology.

[3]  R. Morelos-Zaragoza The art of error correcting coding , 2002 .

[4]  R. Knight,et al.  UniFrac: a New Phylogenetic Method for Comparing Microbial Communities , 2005, Applied and Environmental Microbiology.

[5]  James R. Knight,et al.  Genome sequencing in microfabricated high-density picolitre reactors , 2005, Nature.

[6]  Robert H. Morelos-Zaragoza,et al.  The Art of Error Correcting Coding: Morelos-Zaragoza/The Art of Error Correcting Coding, Second Edition , 2006 .

[7]  Rob Knight,et al.  UniFrac – An online tool for comparing microbial community diversity in a phylogenetic context , 2006, BMC Bioinformatics.

[8]  Philip Hugenholtz,et al.  NAST: a multiple sequence alignment server for comparative analysis of 16S rRNA genes , 2006, Nucleic Acids Res..

[9]  Susan M. Huse,et al.  Microbial diversity in the deep sea and the underexplored “rare biosphere” , 2006, Proceedings of the National Academy of Sciences.

[10]  James A. Foster,et al.  Phylogenetics Clearcut : a fast implementation of relaxed neighbor joining , 2006 .

[11]  Susan M. Huse,et al.  Microbial Population Structures in the Deep Marine Biosphere , 2007, Science.

[12]  F. Bushman,et al.  Short pyrosequencing reads suffice for accurate microbial community analysis , 2007, Nucleic acids research.

[13]  R. Knight,et al.  Global patterns in bacterial diversity , 2007, Proceedings of the National Academy of Sciences.

[14]  F. Bushman,et al.  DNA bar coding and pyrosequencing to identify rare HIV drug resistance mutations , 2007, Nucleic acids research.

[15]  Jonathan P. Bollback,et al.  The Use of Coded PCR Primers Enables High-Throughput Sequencing of Multiple Homolog Amplification Products by 454 Parallel Sequencing , 2007, PloS one.

[16]  Susan M. Huse,et al.  Accuracy and quality of massively parallel DNA pyrosequencing , 2007, Genome Biology.

[17]  G. Casella,et al.  Pyrosequencing enumerates and contrasts soil microbial diversity , 2007, The ISME Journal.

[18]  M. Ronaghi,et al.  A pyrosequencing-tailored nucleotide barcode design unveils opportunities for large-scale sample multiplexing , 2007, Nucleic acids research.