High-speed thermal cycling system and method of use

摘要

A thermal cycling system and method of use are described. The thermal cycling system is based on the-circulation of temperature-controlled water directly to the underside of thin-walled polycarbonate microtiter plates. The water flow is selected from a manifold fed by pumps from heated reservoirs. The plate wells are loaded with typically 15-20 mu l of reagent mix for the PCR process. Heat transfer through the thin polycarbonate is sufficiently rapid that the contents reach thermal equilibrium with the water in less than 15 seconds. Complete PCR amplification runs of 40 three-step cycles have been performed in as little as 14.5 minutes, with the results showing substantially enhanced specificity compared to conventional technology requiring run times in excess of 100 minutes. The plate clamping station is designed to be amenable to robotic loading and unloading of the system. It includes a heated lid, thus eliminating the need for mineral oil overlay of the reactants. The present system includes three or more plate holder stations, fed from common reservoirs but operating with independent switching cycles. The system can be modularly expanded.