Dual polarization interferometry size and density characterisation of DNA immobilisation and hybridisation.
Investigation of nucleic acid interactions was performed using dual polarization interferometry, a novel approach to elucidating molecular interactions. This paper presents a preliminary study of adsorption of single stranded DNA onto functionalised silicon oxynitride, compared with covalent linkage, and avidin-biotin immobilisation. The effect of probe concentration on hybridisation efficiency was also examined. We found that increasing the electrolyte concentration resulted in a decrease of adsorbed DNA and that capture of a biotinylated duplex DNA on an adsorbed avidin layer resulted in four times fewer molecules per cm(2) than for duplex DNA covalently bound via an amine end terminal. The rate of thickness increase of a biotin probe layer on an adsorbed avidin capture layer increased 10-fold when the probe concentration was increased from 0.1 microM to 1 microM. The close grafting density of the higher concentration probe meant that the immobilised probes were unavailable for hybridisation.
Berney
Helen
H
Institute for Nanoscale Science and Technology, Herschel Annex, University of Newcastle upon Tyne, Newcastle upon Tyne, NE1 7RU, UK. hberney@gmail.com
Oliver
Karen
K
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000032:analysis / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D003720:Densitometry / Q000295:instrumentation / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods*; D007368:Interferometry / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012031:Refractometry / Q000295:instrumentation / Q000379:methods
D020099:Coated Materials, Biocompatible; D004247:DNA