Yes, we have tested two methods to digest RNase R-resistant linear RNAs:
- We recommend adding 1 unit of RNase R, 1 unit of XRN-1 (NEB #M0338), and 5 units of RppH (NEB #M0356) to 1 µg of RNA sample in 1X RNase R Reaction Buffer. The reaction should be incubated for 15 or 30 minutes at 37°C for efficient linear RNA removal.
- Pre-treat RNA samples with E. coli Poly(A) Polymerase (NEB #M0276) to lengthen the 3´ ends of the RNAs, which can improve RNase R binding and activity. The RNA needs to be cleaned up before proceeding with the RNase R reaction.
