Rapid melting curve analysis on monolayered beads for high-throughput genotyping of single-nucleotide polymorphisms.
This report describes a rapid solid-phase melting curve analysis method for single-nucleotide polymorphism (SNP) genotyping. The melting curve analysis is based on dynamic allele-specific hybridization (DASH). The DNA duplexes are conjugated on beads that are immobilized on the surface of a microheater chip with integrated heaters and temperature sensors. SNP on PCR products were scored, illustrating the sensitivity and robustness of the system. The method is based on random bead immobilization by microcontact printing. Single-bead detection and multiplexing were performed with a heating rate more than 20 times faster than conventional DASH. Analyses that took more than 15 min could be performed in less that 1 min, enabling ultrarapid SNP analysis. In addition, an array version of the chip was implemented enabling the preparation of an array of bead arrays for high-throughput and rapid SNP genotyping.
Russom
Aman
A
Department of Signals, Sensors and Systems, Microsystem Technology, Royal Institute of Technology, 100 44 Stockholm, Sweden.
Haasl
Sjoerd
S
Brookes
Anthony J
AJ
Andersson
Helene
H
Stemme
Göran
G
D000483:Alleles; D001483:Base Sequence; D004867:Equipment Design; D005838:Genotype; D017403:In Situ Hybridization / Q000379:methods*; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures / Q000379:methods*; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012330:RNA, Double-Stranded / Q000032:analysis* / Q000235:genetics; D013696:Temperature
D012330:RNA, Double-Stranded