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Series GSE3026 Query DataSets for GSE3026
Status Public on Jan 17, 2006
Title Gene expression Profiling of Peripheral Blood Leukocytes after Endotoxin Challenge in Humans
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Peripheral blood mononuclear cells were isolated 0, 30 min, 6 h, 24 h, and 7 day after intravenous endotoxin challenge.
Keywords: time course
 
Overall design Peripheral blood mononuclear cells were isolated 0, 30 min, 6 h, 24 h, and 7 day after intravenous endotoxin challenge using cell preparation tubes (Vacutainer® CPT). The PBMC layer was lysed with RLT buffer, homogenized, and then stored at –80°C. Total RNA was extracted as per RNeasy‚ Midi protocol (Qiagen). Double stranded cDNA was synthesized from total RNA (5 to 20 µg) using 7-d(T)24 primer and SuperScript‰ Double-Stranded cDNA Synthesis Kit and purified by phase lock gel-phenol/chloroform extraction followed by ethanol precipitation. Biotin-labeled cRNA was prepared by in vitro transcription using HighYield‰ RNA Transcript Labeling Kit followed by fragmentation with 5X fragmentation buffer. Fragmented cRNA (10 µg) was hybridized to Affymetrix Hu95Av2 oligonucleotide probe arrays for 16 h at 45∞C. After removal of hybridization fluid, the arrays were washed, stained with streptavidin phycoerythrin (SAPE), and signal amplified by anti-streptavidin antibody using Affymetrix Fluidics Station 400. Probe set signals were measured using Agilent GeneArray Scanner (Affymetrix‚, Santa Clara, CA).
 
Contributor(s) Talwar S, Munson PJ, Barb JJ, Fiuza C, Cintron AP, Logun C, Tropun M, Khan S, Reda D, Shelhamer JH, Danner RL, Suffredini AF
Citation(s) 16403844
Submission date Jul 28, 2005
Last update date Dec 13, 2018
Contact name Anthony F. Suffredini
E-mail(s) asuffredini@cc.nih.gov
Phone 301-496-9320
Fax 301-480-3143
Organization name National Institutes of Health
Department Critical Care Medicine Department
Lab CCMD
Street address Building 10, Rm 7D43, 10 Center Drive
City Bethesda
State/province MD
ZIP/Postal code 20892-1662
Country USA
 
Platforms (1)
GPL8300 [HG_U95Av2] Affymetrix Human Genome U95 Version 2 Array
Samples (73)
GSM66617 n4t0h
GSM66618 n4t30m
GSM66619 n4t6h
Relations
BioProject PRJNA93215

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Supplementary data files not provided

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