biostudies-literatureLeong KWDana-Farber Cancer InstituteNCI NIH HHSNational Institutes of Healthe32https://www.ebi.ac.uk/biostudies/studies/S-EPMC8989544biostudies-literatureUnknown50(6)Detection of low-level DNA mutations can reveal recurrent, hotspot genetic changes of clinical relevance to cancer, prenatal diagnostics, organ transplantation or infectious diseases. However, the high excess of wild-type (WT) alleles, which are concurrently present, often hinders identification of salient genetic changes. Here, we introduce UV-mediated cross-linking minor allele enrichment (UVME), a novel approach that incorporates ultraviolet irradiation (∼365 nm UV) DNA cross-linking either before or during PCR amplification. Oligonucleotide probes matching the WT target sequence and incorporating a UV-sensitive 3-cyanovinylcarbazole nucleoside modification are employed for cross-linking WT DNA. Mismatches formed with mutated alleles reduce DNA binding and UV-mediated cross-linking and Nucleic acids researchMutation enrichment in human DNA samples via UV-mediated cross-linking.PMC8989544R01 CA221874Yu FLeong KWMakrigiorgos GMfalseMutation enrichment in human DNA samples via UV-mediated cross-linking.Detection of low-level DNA mutations can reveal recurrent, hotspot genetic changes of clinical relevance to cancer, prenatal diagnostics, organ transplantation or infectious diseases. However, the high excess of wild-type (WT) alleles, which are concurrently present, often hinders identification of salient genetic changes. Here, we introduce UV-mediated cross-linking minor allele enrichment (UVME), a novel approach that incorporates ultraviolet irradiation (∼365 nm UV) DNA cross-linking either before or during PCR amplification. Oligonucleotide probes matching the WT target sequence and incorporating a UV-sensitive 3-cyanovinylcarbazole nucleoside modification are employed for cross-linking WT DNA. Mismatches formed with mutated alleles reduce DNA binding and UV-mediated cross-linking and 2022-01-01T00:00:00Z2022 Apr2026-03-27T15:18:01.904Z2025-04-04T19:10:47.273ZS-EPMC89895443490467610.1093/nar/gkab1222