title: GSM5701063: wt_cga_mono_r3; Saccharomyces cerevisiae; OTHER
library_construction_protocol: Frozen yeast cultures were lysed mechanically in liquid nitrogen using a freezer mill and thawed into lysis buffer Ribosomes were pelleted through a sucrose cushion then separated into monosomes or disomes by centrifugation through a sucrose gradient and fractionated. Complexes were treated with RNase I and RNA footprints were extracted by phenol/chloroform. Footprint size selection (monosome: 15-35 nt, disome: 40-70 nt) was performed by gel extraction, followed by dephosphorylation, linker ligation, and reverse transcription. Ribosomal cDNA was depleted by subtracted with a pool of biotinylated oligonucleotides. Finally, cDNA was circularized and amplified by PCR with barcode-containing primers.