A Novel Control System for Polymerase Chain Reaction Using a RIKEN GS384 Thermalcycler |
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Authors: | Sasaki Nobuya; Izawa Masaki; Shimojo Masahito; Shibata Kazuhiro; Akiyama Jun-ichi; Itoh Masayoshi; Nagaoka Sumiharu; Carninci Piero; Okazaki Yasushi; Moriuchi Tetsuya; Muramatsu Masami; Watanabe Sachihiko; Hayashizaki Yoshihide |
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Institution: | 1Genome Science Laboratory, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN) 3-1-1 Koyadai, Tsukuba, Ibaraki 305, Japan
2Division of Cell Biology, Cancer Institute, School of Medicine, Hokkaido University Sapporo 060, Japan |
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Abstract: | We have developed a novel high-throughput thermalcycler, theRIKEN GS384, which has a maximum of 1536 wells and whose temperaturecan be controlled accurately and simultaneously for a very smallvolume of a reaction mixture. In practice, the reaction is carriedout using four 384-well (3.5 mm in diameter) plate formats whichcan be automatically moved using a robotic arm. To achieve accuratetemperature control with high thermo-conductivity, we adoptedTeflon-coated aluminum well plates closely sandwiched betweensilicon sheet-covered lids on top and a graphite sheet below.The lids were kept at a higher temperature (2 to 5°C) thanthe reaction wells. The temperature of the 1536 sample wellswas controlled accurately without temperature variability amongthe wells or evaporation, even for samples of very small volume(minimum 2 µl). We also developed a new type of plateformat which is similar to the 384-well place in terms of platesize, shape, and material, but which differs in the number (1536)and size (1.6 mm in diameter) of the wells. Since the amplificationreactions could be done precisely as well, a total of 6144 reactionscan potentially be carried out simultaneously using the GS384thermalcycler. This is very promising for DNA microfabricationtechnology. This thermalcycler offers the advantage of high-throughputDNA analysis which should be useful for DNA diagnoses or forthe human genome project. |
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Keywords: | PCR thermalcycler high-throughput temperature control |
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