[The kit uses total RNA as input, hence no For QuantSeq REV V2 Kits (Cat. No. 225), the recommended input range is 1 - 500 ng of total RNA. Protocol modifications apply for input amounts below 10ng of total RNA (please see the QuantSeq REV V2 User Guide; Appendix B).
No prior poly(A) enrichment or rRNA depletion is required.
The amount of total RNA needed for QuantSeq depends on the poly(A) RNA content of the sample in question. This 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.]
QuantSeq REV is only recommended for input amounts >10 ng of total RNA. If you are limited in terms of input, we recommend using QuantSeq FWD. By choosing longer sequencing read lengths, you are also able to read into the 3' ends.