The For QuantSeq REV V2 Kits (Cat. No. 225), the recommended input range is 10 1 - 500 ng of total RNA. In contrast to QuantSeq FWD, the minimum input is 10 ng Protocol modifications apply for input amounts below 10ng of total RNA (no low input protocol options are available for QuantSeq REVplease see the QuantSeq REV V2 User Guide; Appendix B).
No prior poly(A) enrichment or rRNA depletion is required.
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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.]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.