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Series GSE86337 Query DataSets for GSE86337
Status Public on Sep 02, 2016
Title Transcriptome profiling of 5 human adenocarcinoma cell lines
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Purpose: The aim of this study is to compare the transcriptome profiles of a limited number of lung cancer cell lines with the intention of selecting the two most similar cell lines for a mixture experiment (GSE64098).
Methods - Cell Culture: Five lung adenocarcinoma cell lines (H2228, NCI-H1975, HCC827, H838 and A549) from a range of passages (2-4) were grown on 2 separate occasions in RPMI media (Gibco) supplemented with Glutamax and 10\% fetal calf serum to a 70\% confluence. To replicate common experimental conditions cell lines were treated with 0.01\% Dimethyl sulfoxide (Sigma), which is commonly used as a vehicle in drug treatment experiments. After 6 hours of treatment, cells were collected, snap-frozen on dry ice and stored at -80 degree C until required.
Methods - RNA preparation: Total RNA was extracted from between half a million and million cells using Total RNA Purification Kit (Norgen Biotek) according to the kit instructions. RNA quality and concentration were assessed using Nanodrop and Tapestation RNA ScreenTape (Agilent) respectively.
Methods - RNA-seq: 1 ug of total RNA from each sample were used for RNA-seq library preparation using TruSeq Total Stranded RNA with Ribozero (Illumina) according to manufacturer's instructions. Completed libraries were sequenced on HiSeq 2000 with TruSeq SBS Kit v3- HS reagents (Illumina) as 100 bp single end reads at the Australian Genome Research Facility (AGRF), Melbourne. We obtained on average 28 million for each sample (range from 25 to 29 million). Reads were aligned to the human reference genome hg19 using the Rsubread package (version 1.16.1) (Liao et al. 2013). Single reads were then summarized into gene-level counts using FeatureCounts in the reverse-stranded mode (Liao et al. 2014).
Overall design Total RNA was extracted from lung adenocarcinoma cell lines H2228, NCI-H1975, HCC827, H838 and A549 (2 independent samples for each cell line). Total RNA transcriptome from these samples was profiled by RNA-seq.
Contributor(s) Holik AZ, Ritchie ME
Citation(s) 27899618
Submission date Sep 01, 2016
Last update date May 15, 2019
Contact name Matthew Ritchie
Organization name The Walter and Eliza Hall Institute of Medical Research
Department Epigenetics and Development Division
Street address 1G Royal Parade
City Parkville
State/province Victoria
ZIP/Postal code 3052
Country Australia
Platforms (1)
GPL11154 Illumina HiSeq 2000 (Homo sapiens)
Samples (10)
GSM2300446 A549 rep1
GSM2300447 A549 rep2
GSM2300448 H1975 rep1
BioProject PRJNA341465
SRA SRP083954

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Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE86337_reverse.stranded.unfiltered.count.matrix.txt.gz 356.0 Kb (ftp)(http) TXT
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Processed data are available on Series record
Raw data are available in SRA

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