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J Immunol. 2016 Mar 15;196(6):2902-7. doi: 10.4049/jimmunol.1502563. Epub 2016 Feb 8.

Highly Accurate Sequencing of Full-Length Immune Repertoire Amplicons Using Tn5-Enabled and Molecular Identifier-Guided Amplicon Assembly.

Author information

1
Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064.
2
Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064 vollmers@ucsc.edu.

Abstract

Ab repertoire sequencing is a powerful tool to analyze the adaptive immune system. To sequence entire Ab repertoires, amplicons are created from Ab H chain (IgH) transcripts and sequenced on a high-throughput sequencer. The field of immune repertoire sequencing is growing rapidly and the protocols used are steadily improving; however, thus far, immune repertoire sequencing protocols have not been able to sequence full-length immune repertoires including the entire IgH V region and enough of the IgH C region to identify isotype subtypes. In this study, we present a method that combines Tn5 transposase and molecular identifiers for the highly accurate sequencing of amplicons >500 bp using Illumina short read paired-end sequencing. We then apply this method to Ab H chain amplicons to sequence the first, to our knowledge, highly accurate full-length immune repertoire.

PMID:
26856699
DOI:
10.4049/jimmunol.1502563
[Indexed for MEDLINE]
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