ONT DirectRNA Library preparation for poly(A) estimation
This protocol provides a detailed explanation of of the steps necessary for successful Direct RNA Library preparation for Oxford Nanopore Sequencing. The purpose of this description is to provide optimal conditions for analyses of poly(A) tail length from mRNA using the tailfindr package. The protocol explains the steps needed for RNA sample preparation based on TRIzol extraction and Poly(A) enrichment prior to library preparation. The library preparation protocol is based on protocols provided by Nanopore for RNA-002 kits, yet offers additional advice on what we think is important for a successful library with minimal RNA degradation.