Dicko, Cedric, Hicks, Matthew R., Dafforn, Tim, Vollrath, Fritz, Rodger, Alison and Hoffmann, Søren V.. (2008) Breaking the 200nm limit for routine flow linear dichroism measurements using UV synchrotron radiation. Biophysical Journal, Vol.95 (No.12). pp. 5974-5977. ISSN 0006-3495

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Abstract

The first synchrotron radiation flow linear dichroism spectra are reported. High-quality spectral data can be collected from 450 nm down to 180 nm in contrast to the practical cutoff of not, vert, similar200 nm on benchtop instruments. State-of-the-art microvolume capillary Couette flow linear dichroism was successfully ported to a synchrotron radiation source. The sample volume required is < 50 μL. A characterization of the synchrotron radiation linear dichroism with known DNA and DNA-ligand systems is presented and the viability of the setup confirmed. Typically, wavelengths down to 180 nm are now routinely accessible with a high signal/noise ratio with little limitation from the sample concentration. The 180 nm cutoff is due to the quartz of the Couette cell rather than the beamline itself. We show the application of the simultaneous determination of the sample absorption spectrum to calculate the reduced linear dichroism signal. Spectra for calf thymus DNA, DNA/ethidium bromide, and DNA/4′,6-diamidino-2-phenylindole systems illustrate the quality of data that can be obtained.

Item Type:	Journal Article
Subjects:	Q Science > QD Chemistry
Divisions:	Faculty of Science > Chemistry
Library of Congress Subject Headings (LCSH):	Synchrotron radiation, Linear dichroism, DNA-ligand interactions
Journal or Publication Title:	Biophysical Journal
Publisher:	Biophysical Society
ISSN:	0006-3495
Date:	15 December 2008
Volume:	Vol.95
Number:	No.12
Page Range:	pp. 5974-5977
Identification Number:	10.1529/biophysj.108.139964
Status:	Peer Reviewed
Access rights to Published version:	Open Access
Funder:	Engineering and Physical Sciences Research Council (EPSRC), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Sixth Framework Programme (European Commission) (FP6), Statens naturvidenskabelige forskningsråd (Denmark) [Natural Science Research Council], Carlsbergfondet (Copenhagen, Denmark) [Carlsberg foundation], Lundbeck Foundation, United States. Air Force (USAF)
Grant number:	RII3-CT-2004-506008 (SFP), GR/T09224/01 (EPSRC), EP/E039715/1 (EPSRC), BB/F011199/1 (BBSRC),
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URI:	http://wrap.warwick.ac.uk/id/eprint/917

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