Spectrophotometric analysis of nucleic acids: oxygenation-dependant hyperchromism of DNA |
| |
Authors: | Rupak Doshi Philip J R Day Paolo Carampin Ewan Blanch Ian J Stratford Nicola Tirelli |
| |
Institution: | (1) School of Translational Medicine, Manchester Interdisciplinary Biocentre, The University of Manchester, Manchester, M1 7ND, UK;(2) Faculty of Life Sciences, Manchester Interdisciplinary Biocentre, The University of Manchester, Manchester, M1 7ND, UK;(3) Laboratory of Polymer and Biomaterials, School of Pharmacy & Pharmaceutical Sciences, The University of Manchester, Manchester, M13 9PL, UK;(4) Experimental Oncology, School of Pharmacy & Pharmaceutical Sciences, The University of Manchester, Manchester, M13 9PL, UK;(5) Present address: Department of Pharmacology, School of Biological Sciences, University of Cambridge, Cambridge, CB2 1PD, UK |
| |
Abstract: | The absorbance at 260 nm (A
260) is ubiquitously used for nucleic acid quantification. We show that following oxygenation, DNA solutions experience alterations
in both spectral properties (hyperchromism in the UV region, λ
max 260 nm) and DNA conformation. The spectral changes caused by oxygen–DNA complexation are stable for at least several weeks
at room temperature or several hours at 37 °C, but are also reversible by purging with nitrogen. Our data indicate that DNA
in working solutions might already exist in the oxygen-complexed state, potentially confounding spectrophotometric analyses.
Further, the presence of these complexes does not appear to impart cell toxicity in vitro or affect the biophysical functional
behaviour (e.g. hybridisation) of DNA. Interestingly, our work also suggests that hybridisation could determine a release
of bound oxygen, a phenomenon that could open the way to the use of such systems as oxygen carriers. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|