|
Status |
Public on May 28, 2008 |
Title |
Subject CO, 6h |
Sample type |
RNA |
|
|
Source name |
Human skeletal muscle biopsy from a 3h IL-6 infusion
|
Organism |
Homo sapiens |
Characteristics |
Subject: CO Timepoint: 6h Sex: Male
|
Treatment protocol |
IL-6 infusion from 0h to 3h
|
Extracted molecule |
total RNA |
Extraction protocol |
Trizol, followed by RNEasy (Qiagen) purification of total RNA
|
Label |
biotin
|
Label protocol |
Total RNA was synthesized to double-strand cDNA using Superscript Choice System (Invitrogen, Carlsbad, CA, USA) with an oligo-dT primer containing a T7 RNA polymerase promoter (GenSet, Evry, France). The cDNA was used as a template for in vitro transcription reaction to synthesize biotin-labeled antisense cRNA (BioArray High Yield RNA Transcript Labeling Kit; Enzo Diagnostics, Farmingdale, NY, USA)
|
|
|
Hybridization protocol |
After fragmentation at 94 C for 35 min in fragmentation buffer (40 mM Tris, 30 mM MgOAc, 10 mM KOAc), the labeled cRNA was hybridized for 16 h to Affymetrix HG-U133 Plus 2.0 (Affymetrix Inc., Santa Clara, CA, USA).After hybridization the array was washed and stained with streptavidin phycoerythrin solution using the Fluidics Station 400 (Affymetrix Inc.).
|
Scan protocol |
Finally the arrays were scanned using the GeneChip Scanner 3000 (Affymetrix Inc.) to obtain non-normalized expression levels.
|
Description |
Human skeletal muscle biopsy from vastus lateralis of musculus quadriceps muscle from a male subject.
|
Data processing |
Affymetrix CEL-files were normalized using gcRMA (Wu et al., 2004) available in the Bioconductor package version 1.7 (http://www.bioconductor.org) for the statistical software package R version 2.21 (R Development Core Team, 2005).
|
|
|
Submission date |
Feb 29, 2008 |
Last update date |
Aug 28, 2018 |
Contact name |
Ole Hartvig Mortensen |
E-mail(s) |
[email protected]
|
Organization name |
University of Copenhagen
|
Department |
Department of Biomedical Sciences
|
Lab |
Cellular and Metabolic Research Section
|
Street address |
Panum 6.5.28, Blegdamsvej 3
|
City |
Copenhagen N |
ZIP/Postal code |
2200 |
Country |
Denmark |
|
|
Platform ID |
GPL570 |
Series (1) |
GSE10685 |
Human skeletal muscle biopsies from a 3h IL-6 infusion |
|
Relations |
Reanalyzed by |
GSE119087 |