[Development of high-throughput polymerase chain reaction system and its performance].

摘要

We have developed a high-throughput thermalcycler that allows reduction in reagent volume and large number of PCR amplifications. One microliter of PCR samples can be amplified on the 384-well Teflon-coated aluminum plate. Four plates can be simultaneously subjected to PCR reaction in one instrument (1536 reactions). The heated lid is continually regulated at 2-5 degrees C higher temperature than samples during PCR cycle. This enable to prevent the evaporation in small reaction volume without use of mineral oil. Also, this PCR system employs the special carbongraphite sheet showing high thermal conductivity, resulting in rapid, uniform and reproducible amplification. Furthermore, all system including temperature regulation and open-close of the lids can be automatically controlled by personal computer. The number of the machines which can be synchronously controlled in one integrated PCR system is expanded to achieve the required throughput of PCR reactions. In this paper, we demonstrate that this instrument greatly facilitates to perform a large number of PCR reactions, to reduce the running cost and accelerates the computer-controlled PCR robotization.