Recirculating flow accelerates DNA microarray hybridization in a microfluidic device.
A recirculating microfluidic device fabricated by laminating Mylar and glass was developed for the analysis of hybridization of oligonucleotides to DNA microarrays. The device is part of a system that provides controlled hybridization to DNA probes immobilized in a microarray of polyacrylamide gel pads using recirculation and temperature control. The system was used to obtain real-time kinetics of DNA hybridization and more accurate melting profiles of target-probe duplexes than possible using a static hybridization format. Recirculation shortened the time of perfect match target-probe hybridization from 6 hours to 2 hours and shifted the Td by 1.54 degrees C, relative to static conditions. The experimental results were consistent with a three-dimensional simulation of hybridization using a recirculating buffer system.
Lee
Hyun Ho
HH
Department of Bioengineering, University of Washington, Seattle, WA 98195, USA. hyunho@u.washington.edu
Smoot
James
J
McMurray
Zack
Z
Stahl
David A
DA
Yager
Paul
P
D002021:Buffers; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D005782:Gels; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry*
D002021:Buffers; D005782:Gels; D009841:Oligonucleotides; D004247:DNA