Haplotype phasing of whole human genomes using bead-based barcode partitioning in a single tube View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2017-06-26

AUTHORS

Fan Zhang, Lena Christiansen, Jerushah Thomas, Dmitry Pokholok, Ros Jackson, Natalie Morrell, Yannan Zhao, Melissa Wiley, Emily Welch, Erich Jaeger, Ana Granat, Steven J Norberg, Aaron Halpern, Maria C Rogert, Mostafa Ronaghi, Jay Shendure, Niall Gormley, Kevin L Gunderson, Frank J Steemers

ABSTRACT

Haplotype-resolved genome sequencing promises to unlock a wealth of information in population and medical genetics. However, for the vast majority of genomes sequenced to date, haplotypes have not been determined because of cumbersome haplotyping workflows that require fractions of the genome to be sequenced in a large number of compartments. Here we demonstrate barcode partitioning of long DNA molecules in a single compartment using "on-bead" barcoded tagmentation. The key to the method that we call "contiguity preserving transposition" sequencing on beads (CPTv2-seq) is transposon-mediated transfer of homogenous populations of barcodes from beads to individual long DNA molecules that get fragmented at the same time (tagmentation). These are then processed to sequencing libraries wherein all sequencing reads originating from each long DNA molecule share a common barcode. Single-tube, bulk processing of long DNA molecules with ∼150,000 different barcoded bead types provides a barcode-linked read structure that reveals long-range molecular contiguity. This technology provides a simple, rapid, plate-scalable and automatable route to accurate, haplotype-resolved sequencing, and phasing of structural variants of the genome. More... »

PAGES

852-857

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt.3897

    DOI

    http://dx.doi.org/10.1038/nbt.3897

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1086182235

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/28650462


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