Discussion

Quantitative reverse transcription is a sensitive gene analysis technique which can be used to monitor accumulation of PCR products. PCR amplification results in an increase in target amplicon leading to increased fluorescence from a fluorescent Taqman® probe. Recent advances in the throughput of this technique has enabled its consideration as a screening and hit progression tool within early drug discovery.
Here we describe the development of a high throughput RT-qPCR assay utilising suspension lymphoblasts to enable screening of a 10 thousand compound set. The aim of this screening effort is to identify compounds that reduce messenger RNA levels potentially halting progression of a genetic neurodegenerative disease. Data is nomalised against a housekeeper to determine specificity of the compound ‘effect’. Data will be presented on the assay development, validation and screen, demonstrating the utility of RT-qPCR in a screening environment.