Multiplex amplification of large sets of human exons View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2007-10-14

AUTHORS

Gregory J Porreca, Kun Zhang, Jin Billy Li, Bin Xie, Derek Austin, Sara L Vassallo, Emily M LeProust, Bill J Peck, Christopher J Emig, Fredrik Dahl, Yuan Gao, George M Church, Jay Shendure

ABSTRACT

A new generation of technologies is poised to reduce DNA sequencing costs by several orders of magnitude. But our ability to fully leverage the power of these technologies is crippled by the absence of suitable 'front-end' methods for isolating complex subsets of a mammalian genome at a scale that matches the throughput at which these platforms will routinely operate. We show that targeting oligonucleotides released from programmable microarrays can be used to capture and amplify ∼10,000 human exons in a single multiplex reaction. Additionally, we show integration of this protocol with ultra-high-throughput sequencing for targeted variation discovery. Although the multiplex capture reaction is highly specific, we found that nonuniform capture is a key issue that will need to be resolved by additional optimization. We anticipate that highly multiplexed methods for targeted amplification will enable the comprehensive resequencing of human exons at a fraction of the cost of whole-genome resequencing. More... »

PAGES

931-936

Journal

TITLE

Nature Methods

ISSUE

11

VOLUME

4

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nmeth1110

    DOI

    http://dx.doi.org/10.1038/nmeth1110

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1045927978

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17934468


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