Ribonucleotides embedded within DNA render the DNA sensitive to the formation of single-stranded breaks under alkali conditions. Here, we describe a next-generation sequencing method called hydrolytic end sequencing (HydEn-seq) to map ribonucleotides inserted into the genome of Saccharomyce cerevisiae strains deficient in ribonucleotide excision repair. We use this method to map several genomic features in wild-type and replicase variant yeast strains.
Keywords: Bioinformatics; DNA polymerase; DNA repair; DNA replication; Genomics; HydEn-seq; Next-generation sequencing; RNase H2; Ribonucleotide excision repair.