Hi there,

I am interested in performing polysome profiling with downstream application for RNAseq of monosome and polysome-associated RNAs.

Has anyone had much experience with choice of library preparations for monosome and polysome-associated RNAs?

ribosomal RNA surely is to be more abundant in these samples compared to total RNA extractions? This would lead me towards preferring ribosome-depletion (e.g. riboZero) as opposed to polyA. However PolyA is better for intact fragments and we also intend to sequence total RNA where polyA would be preferred. If we want to compare sequencing between total RNA and mono/poly RNAs it would make sense to use the same library prep?

If you have any experience in choice of library prep (poly A vs ribo-depletion) for polysome-profiling RNAseq experiments please let me know which method you preferred and why

Many thanks,