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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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.
The authors declare no competing financial interests.
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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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