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Scott, Benjamin O. S., Lavesa-Curto, Manuel, Bullard, Desmond R., Butt, Julea N. 
ORCID: https://orcid.org/0000-0002-9624-5226 and Bowater, Richard P. ORCID: https://orcid.org/0000-0002-2745-7807
(2006)
Immobilized DNA hairpins for assay of sequential breaking and joining of DNA backbones.
Analytical Biochemistry, 358 (1).
pp. 90-98.
ISSN 1096-0309
Abstract
Immobilized DNA hairpins are exploited in a novel approach to assay DNA ligases and nucleases. A fundamental characteristic of the assay is that a fluorophore at the remote terminus of the hairpin reports on the integrity of the DNA backbone. The functionality of the protocol is confirmed using ATP- and NAD+-dependent DNA ligases and the nicking enzyme N.BbvCIA. The assay format is amenable to high-throughput analysis and quantitation of enzyme activity, and it is shown to be in excellent agreement with the more laborious electrophoretic approaches that are widely used for such analyses. Significantly, the assay is used to demonstrate sequential breaking and rejoining of a specific nucleic acid. Thus, a simple platform for biochemically innovative studies of pathways in cellular nucleic acid metabolism is demonstrated.
| Item Type: | Article |
|---|---|
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Groups > Biophysical Chemistry (former - to 2017) Faculty of Science > Research Groups > Chemistry of Light and Energy Faculty of Science > Research Groups > Chemistry of Life Processes Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Energy Materials Laboratory |
| Depositing User: | Rachel Smith |
| Date Deposited: | 17 Feb 2011 15:04 |
| Last Modified: | 24 Oct 2022 01:36 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/21408 |
| DOI: | 10.1016/j.ab.2006.08.010 |
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