Development of Novel Hot Start (HS) Reverse Transcriptase Technology

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The Polymerase Chain Reaction (PCR) combined with reverse transcription (RT-PCR and qRT-PCR) is a sensitive method to detect RNA and a crucial technique within bioscience and healthcare research, for example in detecting RNA viruses that exist in very low levels in blood and body fluids of infected individuals. However the sensitivity of the technique can be affected by the enzyme used in the process, Reverse Transcriptase (RTase), forming 'primer dimer' and non specific products at lower temperatures during reaction set up. Our research has shown that the current methods used to improve the sensitivity and specificity of the reaction by inhibiting RTase activity at lower temperatures ('hot start') are inferior and do not inhibit RTase activity sufficiently for ultrasensitive RNA detection. We intend to take an innovative approach to RTase inhibition and develop a novel RTase inhibitor and a range of hot start RTase products for cDNA synthesis, RT-PCR and RT-qPCR to dramatically increase the sensitivity and specificity of RNA detection.