title: Rapid DNA amplification in a capillary tube by natural convection with a single isothermal heater.
abstract: Herein we describe a simple platform for rapid DNA amplification using convection. Capillary convective PCR (CCPCR) heats the bottom of a capillary tube using a dry bath maintained at a fixed temperature of 95°C. The tube is then cooled by the surrounding air, creating a temperature gradient in which a sample can undergo PCR amplification by natural convection through reagent circulation. We demonstrate that altering the melting temperature of the primers relative to the lowest temperature in the tube affects amplification efficiency; adjusting the denaturation temperature of the amplicon relative to the highest temperature in the tube affects maximum amplicon size, with amplicon lengths of ≤500 bp possible. Based on these criteria, we successfully amplified DNA sequences from three different viral genomes in 30 min using CCPCR, with a sensitivity of ~30 copies per reaction.
authors.affiliation: Department of Mechanical Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Wen Pin
authors.initials: WP
authors.lastname: Chou
authors.forename: Ping Hei
authors.initials: PH
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Miao
authors.forename: Long Sheng
authors.initials: LS
authors.lastname: Kuo
authors.forename: Shiou Hwei
authors.initials: SH
authors.lastname: Yeh
authors.forename: Pei Jer
authors.initials: PJ
authors.lastname: Chen
mesh_terms: D018481:Convection; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D016679:Genome, Viral; D006358:Hot Temperature; D019095:Point-of-Care Systems / Q000639:trends; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D017931:DNA Primers; D004247:DNA