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Single-molecule reverse transcription polymerase chain reaction using water-in-oil emulsion

2005; Elsevier BV; Volume: 99; Issue: 3 Linguagem: Inglês

10.1263/jbb.99.293

1389-1723

Michihiko Nakano, Naohito Nakai, Hirofumi Kurita, Jun Komatsu, Kazunori Takashima, Shinji Katsura, Akira Mizuno,

Monoclonal and Polyclonal Antibodies Research

A new method based on a combination of reverse transcription polymerase chain reaction (RT-PCR) and a water-in-oil (W/O) emulsion was developed. Reverse transcription and initial thermal cycles were carried out in droplets of the W/O emulsion. Then, the droplets were united, followed by remaining secondary PCR cycles. This two-step method succeeded in detecting a single RNA molecule.