Create 2024-precise-mapping-single-stranded-DNA-breaks-error-prone-DN…

…A-polymerase.md

content/publication/2024-precise-mapping-single-stranded-DNA-breaks-error-prone-DNA-polymerase.md

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+bibtex_type = "article"
+author="Söderberg O, Wenson L, Heldin J, Martin M, Erbilgin Y, Salman B, Schaal W, Sandbaumhüter F, Jansson E, Chen X, Davidsson A, Stenerlöw B, Spjuth O."
+title="Precise mapping of single-stranded DNA breaks by using an engineered, error-prone DNA polymerase for sequence-templated erroneous end-labelling"
+journal="Research Square"
+year="2024"
+date="2024-10-07T00:00:00+01:55"
+volume=""
+number=""
+preprint = true
+pages=""
+abstract=""
+doi="10.21203/rs.3.rs-5071189/v1"
+url_html="The ability to analyze whether DNA includes lesions is important in identifying mitogenic substances. Until now, the detection of single-stranded DNA breaks (SSBs) has lacked precise methods. To overcome this limitation, we have engineered a chimeric DNA polymerase, Sloppymerase, that is able to replicate DNA in the absence of one nucleotide. In addition to polymerase activity, Sloppymerase demonstrates 5´-3´exonuclease activity. We characterized the activity of Sloppymerase and utilized the enzyme to develop a method for sequence-templated erroneous end-labelling sequencing (STEEL-seq) that is relevant to the mapping of SSBs. Following the omission of a specific nucleotide, e.g., dATP, from the reaction mixture, Sloppymerase introduces mismatches directly downstream of SSBs at positions that should contain deoxyadenosine. The ability to retain sequence information after end-labelling ensures that hits are bona fide SSBs. STEEL-seq works with a variety of sequencing technologies, shown by our successful experiments using Sanger, Illumina, PacBio and Nanopore systems."
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