2025-11-27
ENA first public
ERC000011
ENA-CHECKLIST
University of Cambridge
INSDC center name
public
INSDC status
ERS23954272
SRA accession
E-MTAB-14994:Sample 8
Submitter Id
ArrayExpress
broker name
HeLa
cell line
epithelial cell
cell type
not collected
collection date
human
common name
Protocols: At 5 and 7 hpi, LTM-treated and non-treated cells were flash frozen in an ethanol/dry ice bath and lyzed in the presence of 0.36 mM cycloheximide (CHX). HeLa Ohio cells were grown on 150-mm dishes to a confluency of 80% and infected with CVA-13 at an MOI of 10 to ensure synchronous infection. For lactimidomycin (LTM) treated cells, 30 min before the specified time point, cells were treated with 50 mM LTM for 30 min.  Cells were rinsed with 5 ml of ice-cold PBS, the dishes submerged in a surface of dry ice-ethanol bath for 10 s, transferred to dry ice and 400 µl of lysis buffer [20 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 1% Triton X-100, 100 µg/ml cycloheximide and 25 U/ml TURBO DNase (Life Technologies)] dripped on. Cells were scraped extensively to ensure lysis, collected and triturated with a 26-G needle ten times. Lysates were clarified by centrifugation for 20 min at 13,000 g at 4 ºC, the supernatants recovered and stored in liquid nitrogen. The purification of RPFs was performed using Direct-zol RNA MiniPrep kit (Zymo research). The further methodologies employed were based on the original protocols (Ingolia et al., 2012; Irigoyen et al., 2016), except ribosomal RNA contamination was removed using RiboZero Gold rRNA removal kit (Illumina) and library amplicons were constructed using a small RNA cloning strategy (Guo et al., 2010) adapted to Illumina smallRNA v2 to allow multiplexing. Amplicon libraries were deep sequenced using an Illumina NextSeq platform.
description
adult
developmental stage
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
uterine cervix
organism part
2
replicate
Homo sapiens
scientific_name
Sample 8
title
CVA13
virus