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Target-enrichment strategies for next-generation sequencing

Nature Methods volume 7, pages 111–118 (2010)Cite this article

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An Erratum to this article was published on 01 June 2010

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Abstract

We have not yet reached a point at which routine sequencing of large numbers of whole eukaryotic genomes is feasible, and so it is often necessary to select genomic regions of interest and to enrich these regions before sequencing. There are several enrichment approaches, each with unique advantages and disadvantages. Here we describe our experiences with the leading target-enrichment technologies, the optimizations that we have performed and typical results that can be obtained using each. We also provide detailed protocols for each technology so that end users can find the best compromise between sensitivity, specificity and uniformity for their particular project.

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Figure 1: Approaches to target enrichment.
Figure 2: Suitability of different target-enrichment strategies to different combinations of target size and sample number.
Figure 3: Uniformity of approaches to target enrichment.
Figure 4: Coverage plot for array and solution hybrid capture, for 3.5 Mb of exonic target and whole human exome.
Figure 5: Library prep optimizations for hybrid capture.

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  • 12 April 2010

    In the version of this article initially published, the publication date was incorrectly designated as 28 January 2009 instead of 28 January 2010. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank D. MacArthur, Q. Ayub and C. Tyler-Smith for their work on long PCR and subsequent analyses, P. Akan, A. Palotie, P. Tarpey, H. Arbury and M. Humphries for their work on hybrid capture and E. Sheridan for critical reading of the standard operating procedures. This work was supported by the Wellcome Trust grant WT079643 and by US National Institutes of Health National Human Genome Research Institute grants 5R21HG004749 and 5R01HL094976.

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Authors and Affiliations

  1. The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK

    Lira Mamanova, Alison J Coffey, Carol E Scott, Iwanka Kozarewa, Eleanor Howard & Daniel J Turner

  2. Department of Genome Sciences, University of Washington, Seattle, Washington, USA

    Emily H Turner, Akash Kumar & Jay Shendure

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  1. Lira Mamanova
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Correspondence to Jay Shendure or Daniel J Turner.

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Supplementary Table 1 and Supplementary Protocols 1–6 (PDF 2929 kb)

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Mamanova, L., Coffey, A., Scott, C. et al. Target-enrichment strategies for next-generation sequencing. Nat Methods 7, 111–118 (2010). https://doi.org/10.1038/nmeth.1419

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