{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Mueller SH"],"funding":["Australian Government Research Training Program Scholarship","National Health and Medical Research Council","Australian Research Council"],"pagination":["e5"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9841422"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["51(1)"],"pubmed_abstract":["The activity of enzymes is traditionally characterised through bulk-phase biochemical methods that only report on population averages. Single-molecule methods are advantageous in elucidating kinetic and population heterogeneity but are often complicated, time consuming, and lack statistical power. We present a highly-generalisable and high-throughput single-molecule assay to rapidly characterise proteins involved in DNA metabolism. The assay exclusively relies on changes in total fluorescence intensity of surface-immobilised DNA templates as a result of DNA synthesis, unwinding or digestion. Combined with an automated data-analysis pipeline, our method provides enzymatic activity data of thousands of molecules in less than an hour. We demonstrate our method by characterising three fundamen"],"journal":["Nucleic acids research"],"pubmed_title":["Rapid single-molecule characterisation of enzymes involved in nucleic-acid metabolism."],"pmcid":["PMC9841422"],"funding_grant_id":["DP180100858","DP150100956","FL140100027","2007778"],"pubmed_authors":["Shirbini A","van Oijen AM","Fitschen LJ","Hamdan SM","Spenkelink LM","Mueller SH"],"additional_accession":[]},"is_claimable":false,"name":"Rapid single-molecule characterisation of enzymes involved in nucleic-acid metabolism.","description":"The activity of enzymes is traditionally characterised through bulk-phase biochemical methods that only report on population averages. Single-molecule methods are advantageous in elucidating kinetic and population heterogeneity but are often complicated, time consuming, and lack statistical power. We present a highly-generalisable and high-throughput single-molecule assay to rapidly characterise proteins involved in DNA metabolism. The assay exclusively relies on changes in total fluorescence intensity of surface-immobilised DNA templates as a result of DNA synthesis, unwinding or digestion. Combined with an automated data-analysis pipeline, our method provides enzymatic activity data of thousands of molecules in less than an hour. We demonstrate our method by characterising three fundamen","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Jan","modification":"2026-07-14T15:42:30.955Z","creation":"2025-04-19T12:31:14.555Z"},"accession":"S-EPMC9841422","cross_references":{"pubmed":["36321650"],"doi":["10.1093/nar/gkac949"]}}