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dc.contributor.authorSong, Chenen_US
dc.contributor.authorLiu, Yibinen_US
dc.contributor.authorFontana, Rachelen_US
dc.contributor.authorMakrigiorgos, Alexanderen_US
dc.contributor.authorMamon, Harveyen_US
dc.contributor.authorKulke, Matthew H.en_US
dc.contributor.authorMakrigiorgos, G. Mikeen_US
dc.date.accessioned2016-12-02T15:25:06Z
dc.date.issued2016en_US
dc.identifier.citationSong, Chen, Yibin Liu, Rachel Fontana, Alexander Makrigiorgos, Harvey Mamon, Matthew H. Kulke, and G. Mike Makrigiorgos. 2016. “Elimination of unaltered DNA in mixed clinical samples via nuclease-assisted minor-allele enrichment.” Nucleic Acids Research 44 (19): e146. doi:10.1093/nar/gkw650. http://dx.doi.org/10.1093/nar/gkw650.en
dc.identifier.issn0305-1048en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:29626130
dc.description.abstractPresence of excess unaltered, wild-type (WT) DNA providing no information of biological or clinical value often masks rare alterations containing diagnostic or therapeutic clues in cancer, prenatal diagnosis, infectious diseases or organ transplantation. With the surge of high-throughput technologies there is a growing demand for removing unaltered DNA over large pools-of-sequences. Here we present nuclease-assisted minor-allele enrichment with probe-overlap (NaME-PrO), a single-step approach with broad genome coverage that can remove WT-DNA from numerous sequences simultaneously, prior to genomic analysis. NaME-PrO employs a double-strand-DNA-specific nuclease and overlapping oligonucleotide-probes interrogating WT-DNA targets and guiding nuclease digestion to these sites. Mutation-containing DNA creates probe-DNA mismatches that inhibit digestion, thus subsequent DNA-amplification magnifies DNA-alterations at all selected targets. We demonstrate several-hundred-fold mutation enrichment in diverse human samples on multiple clinically relevant targets including tumor samples and circulating DNA in 50-plex reactions. Enrichment enables routine mutation detection at 0.01% abundance while by adjusting conditions it is possible to sequence mutations down to 0.00003% abundance, or to scan tumor-suppressor genes for rare mutations. NaME-PrO introduces a simple and highly parallel process to remove un-informative DNA sequences and unmask clinically and biologically useful alterations.en
dc.language.isoen_USen
dc.publisherOxford University Pressen
dc.relation.isversionofdoi:10.1093/nar/gkw650en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100565/pdf/en
dash.licenseLAAen_US
dc.titleElimination of unaltered DNA in mixed clinical samples via nuclease-assisted minor-allele enrichmenten
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalNucleic Acids Researchen
dash.depositing.authorSong, Chenen_US
dc.date.available2016-12-02T15:25:06Z
dc.identifier.doi10.1093/nar/gkw650*
dash.authorsorderedfalse
dash.contributor.affiliatedLiu, Yibin
dash.contributor.affiliatedMamon, Harvey
dash.contributor.affiliatedMakrigiorgos, Gerassimos
dash.contributor.affiliatedSong, Chen
dash.contributor.affiliatedKulke, Matthew


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