Temperature and time-resolved total internal reflectance fluorescence analysis of reusable DNA hydrogel chips.
Total internal reflection fluorescence (TIRF) coupled with hydrogel-DNA droplet microarrays covalently bound on PMMA substrates presents a reusable, sensitive platform for evaluating DNA hybridization and for rapid biochip development. Hydrogel microarrays, which contain covalently bound DNA probes, are created via a simple printing and photocross-linking process. TIRF measurements of the arrays display robust reusability, show linear sensitivity down to 5 fmol of fluorescently labeled target DNA, and are sensitive to single basepair mismatches. Additionally, the ability to interrogate larger DNA is shown through studies with PCR amplification hybridization. We conclusively demonstrate an efficient, reproducible, low cost platform for DNA hybridization studies that could be used for fast high-throughput diagnostics as well as biochip development.
Neumann
Thorsten
T
University of Freiburg-IMTEK, Department of Microsystems Engineering, Laboratory for Chemistry and Physics of Interfaces, Georges-Koehler-Allee 103, D-79110 Freiburg, Germany. thoneumann@berkeley.edu
Bonham
Andrew J
AJ
Dame
Gregory
G
Berchtold
Bernd
B
Brandstetter
Thomas
T
Rühe
Jürgen
J
D020137:Base Pair Mismatch; D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors
D015342:DNA Probes; D005456:Fluorescent Dyes; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D004247:DNA