HIGH-SPEED POLYMERASE CHAIN-REACTION IN CONSTANT FLOW

A new simple reactor of the tubing type was developed for polymerase c hain 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 the rmal stages for denaturation, annealing, and elongation of DNA and pri mers as a function of the position in the tube. The amplification yiel d was about a half of that obtained by a commercial thermocycler. More over, the total reaction time from 12 to 18 min, which was one-tenth o f the time generally required by conventional thermocyclers using meta l blocks, assured substantial amplification of a DNA fragment. In addi tion, this reactor could be also used far rapid cycle-sequences. This new device will be easily incorporated into automated and rapid DNA an alysis systems for DNA sequencing.