The recommended input range is 1 - 500 ng of total RNA. Protocol modifications apply for input amounts below 10 ng of total RNA and are outlined in the User guide.
No prior poly(A) enrichment or rRNA depletion of total RNA is required.
Low quality RNA including formalin-fixed parrafin paraffin-embedded (FFPE) samples can also be used for QuantSeqwith QuantSeq following the protocol with modifications for low-quality samples (indicated in the user guide).
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 for further details, or contact support@lexogen.com.