Genotyping by dynamic heating of monolayered beads on a microheated surface
2004 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 25, no 21-22, 3712-3719 p.Article in journal (Refereed) Published
A miniaturized bead-based dynamic allele-specific hybridization (DASH) approach for sing le-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 4degreesC-6degreesC/min compared to earlier reported values of 2degreesC-3degreesC/min, thereby reducing the total analysis time by a factor of 2. Reliable DASH measurement data from areas as small as 12 x 13 mum 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
2004. Vol. 25, no 21-22, 3712-3719 p.
beads, dynamic allele-specific hybridization, microcontact printing, microheater, miniaturization, single-nucleotide polymorphism
IdentifiersURN: urn:nbn:se:kth:diva-5011DOI: 10.1002/elps.200406065ISI: 000225679000021ScopusID: 2-s2.0-10944219819OAI: oai:DiVA.org:kth-5011DiVA: diva2:7465
QC 20100916 201109152005-04-012005-04-012016-03-16Bibliographically approved