The recommended input range is 1 - 500 ng of total RNA.
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.