Illumina MiSeq paired end sequencing
Cells were broken in a French press (SLM Aminco) at 15,000 psi (3 passages) using 10 mL of cell suspension. Ribosomes were purified based on the use of a strong anion exchange monolithic column with an ÄKTA pure FPLC system equipped with fraction collector F9-C to two CIM® QA-0.34 mL monolithic disks. RNA purified from ribosomes was prepared for sequencing using the TruSeq stranded mRNA sample preparation kit (Illumina) according to manufacturer´s instructions, yet without the fragmentation step. The library was quantified via qPCR with the KAPA Library Quantification Kit (Peqlab) and sequenced on a MiSeq desktop sequencer (Illumina), generating paired-end reads with a read length of 75 bp.