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Sample GSM1713694 Query DataSets for GSM1713694
Status Public on Jun 18, 2015
Title IM101_CA04_4d_RNA_3
Sample type RNA
 
Source name Lung_CA04-inoculated_4 days
Organism Mus musculus
Characteristics strain: C57Bl/6J
age: 22 weeks
virus: CA04
time (d.p.i): 4
tissue: lung
biological_replicate: 3
Treatment protocol Mice were anesthetized with isofllurane and inoculated with PBS or PBS containing virus in a volume of 50 μl.
Extracted molecule total RNA
Extraction protocol Total RNA was isolated from TRIzol homogenate of mouse lung.
Label Cy3
Label protocol Total RNA was labeled according to Agilent’s Quick Amp Labeling protocol as part of the Agilent One-Color Microarray-Based Gene Expression Analysis Protocol (Agilent Technology). Total RNA from each sample was linearly amplified and labeled with Cy3-UTP.
 
Hybridization protocol Hybridization was performed according to Agilent One-Color Microarray-Based Gene Expression Analysis Protocol (Agilent Technology). The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000. 1 μg of each labeled cRNA was fragmented by adding 11 μl 10 × Blocking Agent and 2.2 μl of 25×Fragmentation Buffer, then heated at 60 °C for 30 min, and finally 55 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 100μl of hybridization solution was dispensed into the gasket slide and assembled to the gene expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven.
Scan protocol The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).
Description IM101_CA04_4d_3
Data processing Scanned images were analyzed using Agilent Feature Extraction Software (v11.0.1.1). The limma package for R (available on Bioconductor) was used to perform background correction, quantile normalization (normalizeBetweenArrays), and summarization (avereps) to derive a single normalized intensity value per probe. Outlier samples were detected using PCA and by visual inspection of heatmaps, and all data was re-processed after removing outlier samples. All data processing for each of the biological replicates was performed independently of the other.
 
Submission date Jun 17, 2015
Last update date Aug 31, 2016
Contact name Natalie Heller
E-mail(s) natalie.heller@pnnl.gov
Organization name PNNL
Street address 902 Battelle Blvd.
City Richland
ZIP/Postal code 99354
Country USA
 
Platform ID GPL11202
Series (2)
GSE65575 Modeling Host Responses to Understand Severe Human Virus Infections
GSE69945 Mouse lung tissue transcriptome response to H1N1, H5N1, and mutants [mRNA]

Data table header descriptions
ID_REF
VALUE normalized

Data table
ID_REF VALUE
A_55_P1989846 11.01440442
A_55_P1991598 6.845874039
A_55_P2022211 15.61744224
A_55_P1980764 6.778326095
A_55_P1964375 10.77979136
A_51_P128876 16.20821002
A_55_P2121042 6.907373941
A_52_P219230 6.797272878
A_51_P207591 13.42626233
A_55_P2131920 11.37714349
A_55_P2404223 7.184950278
A_55_P2101944 12.90328921
A_52_P358860 6.778326095
A_51_P119031 10.72477257
A_51_P309854 7.644757935
A_51_P343900 10.7383558
A_51_P234359 11.05955295
A_51_P487813 14.77771312
A_52_P613977 11.35739172
A_55_P1957209 10.37697645

Total number of rows: 39429

Table truncated, full table size 978 Kbytes.




Supplementary file Size Download File type/resource
GSM1713694_IM101_CA04_4d_RNA_3.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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