Towards single-molecule DNA sequencing: assays with low nonspecific adsorption.
New DNA sequencing techniques are currently being developed using single-molecule fluorescence-based detection of enzymatic double-strand synthesis. Such application requires surface architectures on which single-stranded templates can be immobilized. A further important attribute is a very low tendency to attract fluorescently labeled bases nonspecifically. On this account, the adsorption behaviour of Cy5-dNTPs on a variety of surface coatings was studied by performing real-time measurements of the DNA synthesis using a supercritical angle fluorescence biosensor. It is demonstrated that polyacrylic acid coatings are an excellent choice to minimize the nonspecific binding of the bases.
Krieg
Alexander
A
Wellcome Trust Centre for Human Genetics, University of Oxford, Headington, Oxford OX3 7BN, UK. alexander.krieg@well.ox.ac.uk
Ruckstuhl
Thomas
T
Seeger
Stefan
S
D000327:Adsorption; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D004247:DNA