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Genotyping by dynamic heating of monolayered beads on a microheated surface
Author(s) -
Russom Aman,
Haasl Sjoerd,
Ohlander Anna,
Mayr Torsten,
Brookes Anthony J.,
Andersson Helene,
Stemme Göran
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406065
Subject(s) - dash , materials science , bead , oligonucleotide , snp genotyping , nanotechnology , analytical chemistry (journal) , genotyping , chromatography , chemistry , computer science , dna , composite material , biochemistry , genotype , gene , operating system
A miniaturized bead‐based dynamic allele‐specific hybridization (DASH) approach for single‐nucleotide polymorphism analysis is presented. Chips with integrated heater and temperature sensors for open‐surface DNA analysis were microfabricated. Microcontact printing using a poly(dimethylsiloxane) (PDMS) stamp was employed to create monolayers of immobilized beads on the surface of the chip. This chip allows fast, well‐controllable temperature ramping. The temperature distribution was homogeneous over the entire heater area. All three possible variants of an SNP site of a synthesized oligonucleotide were accurately scored using the bead‐based DASH approach. Our assay has a nonoptimized temperature ramping rate of 4°C–6°C/min compared to earlier reported values of 2°C–3°C/min, thereby reducing the total analysis time by a factor of 2. Reliable DASH measurement data from areas as small as 12×13 μm was achieved. Our bead‐based DASH approach has enabled a dramatic volume reduction and is a step towards developing a cost‐effective high‐throughput DASH method on arrays of single beads.