Multiplex amplification of large sets of human exons - PubMed (original) (raw)

doi: 10.1038/nmeth1110. Epub 2007 Oct 14.

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

Affiliations

• PMID: 17934468
• DOI: 10.1038/nmeth1110

Multiplex amplification of large sets of human exons

Gregory J Porreca et al. Nat Methods. 2007 Nov.

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 approximately 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.

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Comment in

• Enrichment of super-sized resequencing targets from the human genome.
  Olson M. Olson M. Nat Methods. 2007 Nov;4(11):891-2. doi: 10.1038/nmeth1107-891. Nat Methods. 2007. PMID: 17971778 No abstract available.

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