Genotyping by dynamic heating of monolayered beads on a microheated surface
2004 (English)In: Micro Total Analysis Systems 2004: Proceedings of [Mu] TAS 2004 8th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Malmö, Sweden, September 26-30, 2004, Royal Society of Chemistry, 2004, Vol. 2, 303-305 p.Conference paper (Refereed)
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.
Place, publisher, year, edition, pages
Royal Society of Chemistry, 2004. Vol. 2, 303-305 p.
Beads, Dynamic allele-specific hybridization, Microcontact printing, Microheater, Miniaturization, Single-nucleotide polymorphism
IdentifiersURN: urn:nbn:se:kth:diva-91801ISBN: 9780854046430OAI: oai:DiVA.org:kth-91801DiVA: diva2:784942
8th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Malmö, Sweden, September 26-30, 2004
QC 201502092015-01-312012-03-202015-02-09Bibliographically approved