High speed polymerase chain reaction in constant flow.
A new simple reactor of the tubing type was developed for polymerase chain reaction (PCR). A thin Teflon capillary tube was used as a tubing reactor in which the reaction mixture of PCR was driven by a pump at a constant flow rate. The sample was treated with three successive thermal stages for denaturation, annealing, and elongation of DNA and primers as a function of the position in the tube. The amplification yield was about a half of that obtained by a commercial thermocycler. Moreover, the total reaction time from 12 to 18 min, which was one-tenth of the time generally required by conventional thermocyclers using metal blocks, assured substaintial amplification of a DNA fragment. In addition, this reactor could be also used for rapid cycle-sequences. This new device will be easily incorporated into automated and rapid DNA analysis systems for DNA sequencing.
Nakano
H
H
Department of Applied Biological Sciences, School of Agriculture, Nagoya University, Japan.
Matsuda
K
K
Yohda
M
M
Nagamune
T
T
Endo
I
I
Yamane
T
T
D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods; D013698:Templates, Genetic; D013997:Time Factors
D004269:DNA, Bacterial