Chemical gradient-mediated melting curve analysis for genotyping of SNPs
2009 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 30, no 14, 2536-2543 p.Article in journal (Refereed) Published
This report describes a microfluidic solid-phase chemical gradient-mediated melting curve analysis method for SNP analysis. The method is based on allele-specific denaturation to discriminate mismatched (MM) from perfectly matched (PM) DNA duplexes upon exposure to linear chemical gradient. PM and MM DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device designed with dams, keeping the beads trapped perpendicular to a gradient generating channel. Two denaturants, formamide and urea, were tested for their ability to destabilize the DNA duplex by competing with Watson-Crick pairing. Upon exposure to the chemical gradient, rapid denaturing profile was monitored in real time using fluorescence microscopy. The results show that the two duplexes exhibit different kinetics of denaturation profiles, enabling discrimination of MM from PM DNA duplexes to score SNP.
Place, publisher, year, edition, pages
2009. Vol. 30, no 14, 2536-2543 p.
Chemical gradient, Formamide, Melting curve analysis, SNP, Urea
IdentifiersURN: urn:nbn:se:kth:diva-89922DOI: 10.1002/elps.200800729ISI: 000269041500017OAI: oai:DiVA.org:kth-89922DiVA: diva2:503994
QC 201203122012-02-172012-02-172012-03-12Bibliographically approved