Translation regulatory divergence in yeast
Strand-specific Ribosome profiling (Ribo-seq) library constructed as described in Ingolia et al., Cell 147 2011 789-802. Polysome associated mRNA was digested with RNaseI and monosomes were purified by pelleting through a sucrose cushion. Ribosome protected fragments were  gel purified and ligated to a 3' adapter, and reverse-transcribed.  Resulting cDNA was circularized and contaminating rRNA sequences were removed by subtractive hybridization prior to 12 cycles of PCR to incorporate Illumina library sequences and barcodes.