title: GSM3146282: ribosome footprints_ltm_ifn_vir; Homo sapiens; OTHER
library_construction_protocol: Samples were flash frozen in liquid nitrogen and lysed. A portion of the lysate was used to generate RNA-Seq libraries using NEBNext 893 Ultra Directional RNA Library Prep Kit  after depleting ribosome RNA using the Ribozero Gold kit. For ribosome profiling, we performed nuclease footprinting treatment by adding RNase I . We collected ribosome protected mRNA fragments using MicroSpin S-400 HR Columns, and purified RNA from the flow through for size selection. We gel-purified ribosome protected fragments with length between 26 and 34 nt using RNA oligo size markers. We used polyA tailing instead of linker ligation, performed reverse transcription, circularization of cDNA, and PCR amplification of cDNA. Libraries were sequenced on Illumina HiSeq 2500 in 50bp single end mode.
attributes.tag: GEO Accession
attributes.value: GSM3146282