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[高通量聚合酶链反应系统的研制及其性能]

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

作者信息

Sasaki N

机构信息

Division of Cell Biology, Hokkaido University School of Medicine, Sapporo, Japan.

出版信息

Hokkaido Igaku Zasshi. 1997 May;72(3):249-59.

PMID:9226465
Abstract

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.

摘要

我们开发了一种高通量热循环仪,它能够减少试剂用量并进行大量的PCR扩增。一微升PCR样品可在涂有特氟龙的384孔铝板上进行扩增。一台仪器可同时对四块板进行PCR反应(1536个反应)。在PCR循环过程中,加热盖的温度比样品持续高2 - 5摄氏度进行调节。这使得在不使用矿物油的情况下,能防止小反应体积中的蒸发。此外,该PCR系统采用了具有高导热性的特殊碳石墨片,从而实现快速、均匀且可重复的扩增。再者,包括温度调节和盖子开合在内的所有系统都可由个人计算机自动控制。一个集成PCR系统中可同步控制的机器数量得以扩展,以实现所需的PCR反应通量。在本文中,我们证明该仪器极大地便于进行大量的PCR反应、降低运行成本并加速计算机控制的PCR自动化。

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