Single-molecule reverse transcription polymerase chain reaction using water-in-oil emulsion.
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.
Nakano
Michihiko
M
Department of Ecological Engineering, Toyohashi University of Technology, 1-1 Hibarigaoka, Tempaku-cho, Toyohashi, Aichi 441-8580, Japan.
Nakai
Naohito
N
Kurita
Hirofumi
H
Komatsu
Jun
J
Takashima
Kazunori
K
Katsura
Shinji
S
Mizuno
Akira
A
D005591:Chemical Fractionation / Q000379:methods; D004655:Emulsions / Q000737:chemistry; D006678:HIV / Q000235:genetics* / Q000302:isolation & purification*; D009821:Oils / Q000737:chemistry*; D012367:RNA, Viral / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014867:Water / Q000737:chemistry*
D004655:Emulsions; D009821:Oils; D012367:RNA, Viral; D014867:Water