GSM1568346: vps35 tif; Saccharomyces cerevisiae; ncRNA-Seq
Cells were lysed using a cryogenic mixer mill. Lysates were clarified by centrifugation, and digested using Rnase I. Ribosomes were isolated by ultracentrifugation through a sucrose cushion, and RNA was extracted from the pellet. RNA fragments of approximately 30 nucleotides were extracted from a polyacrylamide gel. A DNA adapter was ligated to the 3' of fragments and used to prime reverse transcription. cDNA were circularized and rRNA-originating fragments were removed using biotinylated subtraction oligonucleotides. The library was barcoded during PCR amplification.