WT and H2O2 treated yeast ribosomal RNA input and translating fractions
500 ng total of polyA tailed yeast total RNA were individually ligated to pre-annealed custom RT adaptors (IDT)  in four separate eppendorfs, using concentrated T4 DNA Ligase (NEB-M0202T), and were reverse transcribed using SuperScript III Reverse Transcriptase (Thermo Fisher Scientific-18080044). The products were purified using 1.8X Agencourt RNAClean XP beads (Fisher Scientific-NC0068576), washing with 70% freshly prepared ethanol. In total, 50 ng of reverse transcribed RNA from each reaction was pooled, and RNA Adapter (RMX), composed of sequencing adapters with motor protein, was ligated onto the RNA:DNA hybrid. The mix was purified using 1X Agencourt RNAClean XP beads, washing with Wash Buffer twice. The sample was then eluted in Elution Buffer and mixed with RNA Running Buffer prior to loading onto a primed R9.4.1 flow cell