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一种使用理化学研究所GS384型热循环仪的新型聚合酶链式反应控制系统。

A novel control system for polymerase chain reaction using a RIKEN GS384 thermalcycler.

作者信息

Sasaki N, Izawa M, Shimojo M, Shibata K, Akiyama J, Itoh M, Nagaoka S, Carninci P, Okazaki Y, Moriuchi T, Muramatsu M, Watanabe S, Hayashizaki Y

机构信息

Genome Science Laboratory, Tsukuba Life Science Center, Ibaraki, Japan.

出版信息

DNA Res. 1997 Dec 31;4(6):387-91. doi: 10.1093/dnares/4.6.387.

Abstract

We have developed a novel high-throughput thermalcycler, the RIKEN GS384, which has a maximum of 1536 wells and whose temperature can be controlled accurately and simultaneously for a very small volume of a reaction mixture. In practice, the reaction is carried out using four 384-well (3.5 mm in diameter) plate formats which can be automatically moved using a robotic arm. To achieve accurate temperature control with high thermo-conductivity, we adopted Teflon-coated aluminum well plates closely sandwiched between silicon sheet-covered lids on top and a graphite sheet below. The lids were kept at a higher temperature (2 to 5 degrees C) than the reaction wells. The temperature of the 1536 sample wells was controlled accurately without temperature variability among the wells or evaporation, even for samples of very small volume (minimum 2 microliters). We also developed a new type of plate format which is similar to the 384-well place in terms of plate size, shape, and material, but which differs in the number (1536) and size (1.6 mm in diameter) of the wells. Since the amplification reactions could be done precisely as well, a total of 6144 reactions can potentially be carried out simultaneously using the GS384 thermalcycler. This is very promising for DNA microfabrication technology. This thermalcycler offers the advantage of high-throughput DNA analysis which should be useful for DNA diagnoses or for the human genome project.

摘要

我们开发了一种新型高通量热循环仪——理研GS384,它最多有1536个孔,能够对极少量反应混合物精确且同时地进行温度控制。实际上,反应是使用四种384孔(直径3.5毫米)板形式进行的,这些板可以通过机械臂自动移动。为了通过高导热性实现精确的温度控制,我们采用了涂有特氟龙的铝制孔板,其被紧密夹在顶部覆盖有硅片的盖子和下面的石墨片之间。盖子的温度保持得比反应孔高(2至5摄氏度)。即使对于极少量(最小2微升)的样品,1536个样品孔的温度也能被精确控制,孔与孔之间没有温度变化,也没有蒸发现象。我们还开发了一种新型板形式,其在板的尺寸、形状和材料方面与384孔板相似,但孔的数量(1536个)和尺寸(直径1.6毫米)不同。由于扩增反应也能精确进行,使用GS384热循环仪总共可以潜在地同时进行6144个反应。这对于DNA微制造技术非常有前景。这种热循环仪具有高通量DNA分析的优势,这对于DNA诊断或人类基因组计划应该是有用的。

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