What is the recommended amount of input template for each RT² qPCR Primer Assay?

The useful range of input total RNA for the first strand cDNA template synthesis (reverse transcription) reaction is between 100ng and 5 µg. For initial experiments, we recommend using between 0.5 to 1 µg of input total RNA, and using 1 µl of either undiluted template or template pre-diluted 1:10 for each 25-µl RT² qPCR Assay reaction.