title: Practical innovations for high-throughput amplicon sequencing.
abstract: We describe improvements for sequencing 16S ribosomal RNA (rRNA) amplicons, a cornerstone technique in metagenomics. Through unique tagging of template molecules before PCR, amplicon sequences can be mapped to their original templates to correct amplification bias and sequencing error with software we provide. PCR clamps block amplification of contaminating sequences from a eukaryotic host, thereby substantially enriching microbial sequences without introducing bias.
authors.affiliation: 1] Department of Biology, University of North Carolina, Chapel Hill, North Carolina, USA. [2] Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina, USA. [3].
authors.forename: Derek S
authors.initials: DS
authors.lastname: Lundberg
authors.forename: Scott
authors.initials: S
authors.lastname: Yourstone
authors.forename: Piotr
authors.initials: P
authors.lastname: Mieczkowski
authors.forename: Corbin D
authors.initials: CD
authors.lastname: Jones
authors.forename: Jeffery L
authors.initials: JL
authors.lastname: Dangl
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics; D002965:Classification / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D054892:Metagenome; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012984:Software; D012988:Soil Microbiology
chemical_list: D012336:RNA, Ribosomal, 16S