Detection technique of asymmetric RT-PCR-based amplified single-stranded DNA and its application to biosensor for detection of mRNA for cyanobacteria, Anabaena variabilis

A real-time detection technique for mRNA was developed using a specific DNA probe. The design of the probe was based on a sequence polymorphism within the expressed mRNA of Anabaena variabilis PCC 7120 by using the BIAcore 2000 biosensor, which uses surface plasmon resonance (SPR). The reverse transcription polymerase chain reaction (RT-PCR)-amplified single-stranded DNA (ssDNA), specific for ribonuclease P RNA B (rnpB) from A. variabilis, was determined by an SPR biosensor. Through these detection methods, the ssDNA amplified from rnpB could be detected within 10 min after asymmetric RT-PCR.