Instrument: Illumina HiSeq 2000
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: SINGLE
Construction protocol: The cells were harvested by aspiration of media and immediate addition of ice-cold PBS containing 100 µg/mL cycloheximide. The PBS plus cycloheximide was aspirated with a vacuum aspirator and 400 µL lysis buffer (20 mM Tris-Cl pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM dithiothreitol, 100 µg/mL cycloheximide, 1% Triton X-100 and 25 U/mL DNase I (Promega)) was added. Cells were lysed and ribosome footprint or fragmented RNA libraries were generated as described in Ingolia et al. Curr Protoc Mol Biol Chapter 4, Unit 4 18 (2013).