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SRX17314773: GSM6523409: Ribo-seq MCMV 2hpi - Harringtonin - Rep 3; Mus musculus; OTHER
3 ILLUMINA (Illumina HiSeq 2000) runs: 25.6M spots, 1.3G bases, 953.8Mb downloads

External Id: GSM6523409_r1
Submitted by: Institute of Virology and Immunobiology, University of Wuerzburg
Study: Ribosome profiling of MCMV
show Abstracthide Abstract
Swiss murine embryonic fibroblasts (NIH-3T3) were infected with wild-type Smith strain mouse cytomegalovirus (MCMV) at a multiplicity of infection (MOI) of 10. Ribosome profiling was performed at various times during infection with minor modification to the protocol described in Stern-Ginossar N et al., Science 2012. The detailed protocol is described in Rutkowski et al, Nature commun 2015, and Whisnant et al., Nature commun. 2020. Translation start site profiling was performed by culturing cells in presence of Harringtonin or Lactimidomycin for 30 min prior to cell harvest. Overall design: Ribosome profiling was performed a 0, 2, 6, 12, 24 and 48 h post infection. Translation start site profiling was performed using Harringtonin (2µg/ml for for 30 min) or Lactimidomycin (50µM for 30 min) prior to cell lysis. Two biological replicates were analysed.
Sample: Ribo-seq MCMV 2hpi - Harringtonin - Rep 3
SAMN30571938 • SRS14876261 • All experiments • All runs
Organism: Mus musculus
Library:
Name: GSM6523409
Instrument: Illumina HiSeq 2000
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: SINGLE
Construction protocol: Cells were snap frozen followed by cell lysis in presence of cycloheximide. Following removal of cell nuclei by centrifugation and stringent RNAse digest, ribosomes were pelleted through a sucrose cushion. Translated RNA fragments were recovered by size selection. Translated RNA fragments were cloned following the protocol described by Stern-Ginossar N et al. Science 2012. rRNA depletion involved additional biotinylated oligos to improve depletion. To efficiently remove all biotinylated oligos complementary to specific rRNA sequences, two rounds of streptavidin depletion were performed. cDNA libraries were amplified by PCR and sequenced (HiSeq 50nt reads).
Runs: 3 runs, 25.6M spots, 1.3G bases, 953.8Mb
Run# of Spots# of BasesSizePublished
SRR2130820625,534,0731.3G949.8Mb2022-12-30
SRR2130820738,8841.9M2.2Mb2022-12-30
SRR2130820838,1561.9M1.8Mb2022-12-30

ID:
24135714

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