Please cite this article in press as: Han, J., et al., Discovery of novel non-synonymous SNP variants in 988 candidate genes from 6 centenarians by target capture and next-generation sequencing. Mech. Ageing Dev. (2013), http://dx.doi.org/10.1016/ J. Han et al./Mechanisms of Ageing and Development xxx (201analysis of 6 centenarians revealed a total of 7027 genetic variations in 988 candidate genes, excluding variations annotated as ncRNAs. Among these, we identified 89 novel nsSNPs, of which 67 were assayed for validation and genotyping by iPLEX MassArray method. We found that these novel nsSNPs were rare in our populations. Nevertheless, we validated 51 novel nsSNPs (76%) as true positives. Our customized target capture and next-generation sequencing can reproducibly detect genetic variation in candidate genes. This licensing for a large-scale genetic discovery study to identify all possible variations, including rare variants, in a cost-effective manner. We hope to identify regulatory regions, including 2 kb proximal promoter, exonintron junctions, and 50 and 30 UTRs since our targets contain them. We detected additional centenarians harboring the p.P391L variant in ZMIZ1 or the p.L40F variant in PLCD4 that were predicted to be damaging or deleterious by SIFT and PolyPhen2 analysis. There were also 11 variants detected only in the discovery panel of 6 centenarians. Interestingly, all but the p.I342V variantin USP24 were predicted to be functional by the SIFT or PolyPhen2 analysis. These variants could be good candidates for further genotyping analysis in a larger sample size and for functional analysis to assess the relevance of the genetic variations to longevity.Association study identifies a single major locus contributing to survival into old age: the APOE locus revisited. 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Dristo M.A., Banks E., Poplin R., Garimella K.V., Maguire J.R., Hartl C. Philippakis A.A. del Angel G. Rivas M.A. Hanna M. Menna A. Fennell T.J. Kernytsky A.M. Sivachenko A.Y. Cibulskis K. Gabriel S.B. Altshuler D. Daly M.J. (2013), A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nature Genetics.

Garinis G.A., van der Horst G.T.J,. Vijg J. and Hoeijmakers J.H.J.(2008), DNA damage and ageing: new-age ideas for an age-old problem. Nature Cell Biology.