The recommended input range is 1 - 500 ng of total RNA. Protocol modifications apply for input amounts below 10 ng of total RNA, see: What is the minimum input amount for QuantSeq FWD? Are there any protocol modifications for low input amounts?
No prior poly(A) enrichment or rRNA depletion of total RNA is required.
Low quality RNA including formalin-fixed parrafin embedded (FFPE) samples can also be used for QuantSeq.
The ideal amount of total RNA needed for QuantSeq depends on the poly(A) RNA content of the sample in question. The protocol was tested extensively with various cell cultures, animal and plant tissues, yeast, fungi, drosophila, and human reference RNA.
As a starting point, we recommend performing the protocol initially with 500 ng total RNA. RNA inputs ≥200 ng are recommended to detect low abundant transcripts efficiently.
NOTE: Input amounts <1 ng can also work, please consult the QuantSeq User Guides (015UG009, and 113UG227) for further details, or contact support@lexogen.com.