Automated high-throughput RT-PCR v2

This protocol provides an outline for planning, setting up, and running an epMotion® M5073 for high-throughput RT-PCR. This protocol leverages a set of pre-written template programs to accommodate different numbers of gene expression targets. The template programs have been designed to dispense 2 μL of diluted RNA and 8 μL of one-step master mix (including primers and probes) to create 10 μL reactions on a 384-well PCR plate. Diluted RNA is to be arranged in a 96-well PCR plate for efficiency. We also provide the steps necessary to create custom RT-PCR runs for more advanced users. These steps also detail the settings used in our predefined programs. Users can expect consistent results with minimal variability between technical replicates that are quick and reliable.