| Budget Amount *help |
¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000)
Fiscal Year 2014: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2013: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2012: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
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| Outline of Final Research Achievements |
In this study, we aimed to develop a technique for digital counting of low copy viral DNA within infected cells. We developed a microfluidic device for generation of microdroplets for droplet-based digital PCR. In this method, the target genes were separately amplified within picoliter droplets and detected with fluorescent microscopy. The number of fluorescent droplets correlated with the copy numbers of target genes. In addition, we constructed lentivirus vector modeled after HIV. After optimization of the reaction conditions of droplet digital PCR, we demonstrated its highly sensitive detection of proviral DNA within a small number of infected cells during early infection process.
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