Comparison of ribosome footprints produced with Benzonase and RNAse I
In order to introduce protocol variants in the Ribo-seq approach, we decided to use Benzonase to digest the ribosomes since it produces footprints that are ready to proceed with library preparation. On the other hand, the most used RNAse for footprint production, RNAse I, involves further steps of end repair and re-purification of RNA. To validate our approach, we generate equivalent amounts of footprints with both enzymes and proceed with sequencing to compare each enzyme's footprint in terms of size, mapping distribution and periodicity, among other features, and to contrast the translatome quantification derived by each enzyme.