FAQ: What if I have already fragmented my total RNA and need to perform the First Strand Synthesis Reaction?

If you are starting from fragmented total RNA, we recommend using the NEBNext rRNA Depletion Kit (Human/Mouse/Rat; #E6310) prior to first strand synthesis. Follow the protocol in Chapter 3 of the Ultra II Directional RNA Library Prep Kit (#E7760) for strand-specific RNA-seq workflows. For non-directional RNA-seq workflows, follow the protocol in Chapter 3 of the Ultra II RNA Library Prep Kit (#E7770).