A simple device using magnetic transportation for droplet-based PCR |
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Authors: | Tetsuo Ohashi Hiroki Kuyama Nobuhiro Hanafusa Yoshiyuki Togawa |
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Institution: | (1) Life Science Laboratory, Analytical and Measuring Instruments Division, Shimadzu Corporation, Kyoto 604-8511, Japan;(2) Institute for Protein Research, Osaka University, Suita 565-0871, Osaka, Japan |
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Abstract: | The Polymerase chain reaction (PCR) was successfully and rapidly performed in a simple reaction device devoid of channels,
pumps, valves, or other control elements used in conventional lab-on-a-chip technology. The basic concept of this device is
the transportation of aqueous droplets containing hydrophilic magnetic beads in a flat-bottomed, tray-type reactor filled
with silicone oil. The whole droplets sink to the bottom of the reactor because their specific gravity is greater than that
of the silicone oil used here. The droplets follow the movement of a magnet located underneath the reactor. The notable advantage
of the droplet-based PCR is the ability to switch rapidly the proposed reaction temperature by moving the droplets to the
required temperature zones in the temperature gradient. The droplet-based reciprocative thermal cycling was performed by moving
the droplets composed of PCR reaction mixture to the designated temperature zones on a linear temperature gradient from 50°C
to 94°C generated on the flat bottom plate of the tray reactor. Using human-derived DNA containing the mitochondria genes
as the amplification targets, the droplet-based PCR with magnetic reciprocative thermal cycling successfully provided the
five PCR products ranging from 126 to 1,219 bp in 11 min with 30 cycles. More remarkably, the human genomic gene amplification
targeting glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was accomplished rapidly in 3.6 min with 40 cycles. |
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Keywords: | Magnetic beads Droplets Polymerase chain reaction Microfluidic technology |
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