5'-End Mapping in Human Mitochondrial DNA

Andranik Durgaryan; Anders R Clausen

doi:10.1007/978-1-0716-2922-2_22

5'-End Mapping in Human Mitochondrial DNA

Methods Mol Biol. 2023:2615:315-325. doi: 10.1007/978-1-0716-2922-2_22.

Authors

Andranik Durgaryan¹, Anders R Clausen²

Affiliations

¹ Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.
² Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden. anders.clausen@gu.se.

PMID: 36807801
DOI: 10.1007/978-1-0716-2922-2_22

Abstract

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.

MeSH terms

DNA Replication
DNA, Mitochondrial* / genetics
High-Throughput Nucleotide Sequencing
Humans
Mitochondria* / genetics
Sequence Analysis, DNA

Substances

DNA, Mitochondrial