Single-nucleotide polymorphism analysis by allele-specific extension of fluorescently labeled nucleotides in a microfluidic flow-through device
2003 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 24, no 1-2, 158-161 p.Article in journal (Refereed) Published
We describe a microfluidic approach for allele-specific extension of fluorescently labeled nucleotides for scoring of single-nucleotide polymorphism (SNP). The method takes advantage of the fact that the reaction kinetics differs between matched and mismatched configurations of allele-specific primers hybridized to DNA template. A microfluidic flow-through device for biochemical reactions on beads was used to take advantage of the reaction kinetics to increase the sequence specificity of the DNA polymerase, discriminating mismatched configurations from matched. The volume of the reaction chamber was 12.5 nL. All three possible variants of an SNP site at codon 72 of the p53 gene were scored using our approach. This work demonstrates the possibility of scoring SNP by allele-specific extension of fluorescently labeled nucleotides in a microfluidic flow-through device, The sensitive detection system and easy microfabrication of the microfluidic device enable further miniaturization and production of an array format of microfluidic devices for high-throughput SNP analysis.
Place, publisher, year, edition, pages
2003. Vol. 24, no 1-2, 158-161 p.
filter chamber, flow-through, fluorescence, microfluidics, single-nucleotide polymorphism
IdentifiersURN: urn:nbn:se:kth:diva-5008DOI: 10.1002/elps.200390008ISI: 000180637300021PubMedID: 12652586OAI: oai:DiVA.org:kth-5008DiVA: diva2:7462
QC 20100929 NR 201408052005-04-012005-04-012012-02-17Bibliographically approved