Discussion

AZ employs a large number of different techniques for high throughput screening and hit generation and validation including standard luminescence, absorption and reporter gene assays as well as complex high content biology assays and label free technologies. Traditionally, quantitative PCR (qPCR) has been used for molecular profiling and target identification but not in the context of high-throughput screening. Owing to limitations on workflow automation, cost of reagents and/or miniaturisation opportunities. . Recent advances have moved Reverse Transcription qPCR (RT-qPCR) forward such as improvements in liquid handling, the launch of higher throughput platforms and the release of one-step products. Such one-step reagents enable the user to go straight from a cellular assay format to qPCR without the need for cumbersome multi-step RNA purification protocols. This presentation describes the use of a One-Step accelerated workflow using lysates generated by the Realtime Ready Cell Lysis kit in downstream quantitative RT-qPCR. Cells can be cultured, lysed, and transferred from acoustically qualified, tissue culture treated microplates using the Labcyte Echo, into a 384- or 1536-well PCR plate. The miniaturized and high throughput workflow enables future application of this sensitive assay technology, with particular impact against phenotypic assays and those using rare cell types.