Are there recommendations for low RNA input?

RNA input amounts lower than 100 ng (for total RNA or enriched small RNA) require protocol adjustments. These adjustments include dilution of the 3' Adapter (A3), 5' Adapter (A5) and Reverse Transcription Primer (RTP) and adjusted endpoint PCR cycle numbers (see Appendix B of the User Guide).

The minimum amount of total RNA input depends on the small RNA content of the sample in question.

For samples with a small RNA content of 10 %, 1 ng total RNA may be used as the minimum input.

Increase the input material for total RNA samples with lower small RNA content.