Here, we describe an assay that enables mapping of 5'-ends across the genome using next-generation sequencing on an Illumina platform, 5'-End-sequencing (5'-End-seq). We use this method to map free 5'-ends in mtDNA isolated from fibroblasts. This method can be used to answer key questions regarding DNA integrity, DNA replication mechanisms and to identify priming events, primer processing, nick processing, and double strand break processing on the entire genome.
Keywords: 5′-End-seq; Bioinformatics; DNA polymerase; DNA replication; Genomics; Next-generation sequencing; RNase H1; mtDNA.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.