|
| Status |
Public on Mar 06, 2007 |
| Title |
Neurosphere-like glioma Cell Line R18 |
| Sample type |
RNA |
| |
|
| Source name |
Neurosphere-like human glioblastoma
|
| Organism |
Homo sapiens |
| Characteristics |
Driven by CD133+ cancer stem cells
|
| Growth protocol |
DMEM-F12 Medium supplemented with EGF, FGF, LIF and B27
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Quiagen RNAeasy according to manufactors protocoll
|
| Label |
biotin
|
| Label protocol |
In detail, 2,5 µg of total RNA was converted to cDNA using a T7-oligo-d(T)24 primer and SuperScript II reverse transcriptase. Second strand cDNA synthesis and blunt ending was performed using RNase H, E.coli DNA polymerase I, E.coli DNA ligase and T4 DNA Polymerase. Following cleanup, cDNA was used for in vitro transcription amplification reaction using T7 RNA Polymerase and a biotinylated nucleotide analog/ribonucleotide mix to produce biotinylated cRNA.
|
| |
|
| Hybridization protocol |
Thereafter, the labeled cRNA was purified and fragmented by incubation at 94°C for 35 min. The quality of cRNA and the fragmentation were checked by microcapillary electrophoresis on an Agilent 2100 bioanalyzer, and 15 µg of biotinylated cRNA were hybridized to HG-U133plus 2.0 GeneChip® arrays for 16 h at 45°C with constant rotation at 60 rpm. All microarrays were processed in an Affymetrix GeneChip® Fluidics Station 400. Following staining with streptavidin R-phycoerythrin conjugate and washing cycles,
|
| Scan protocol |
The microarrays were scanned using the GeneChip® Scanner 3000 7G.
|
| Description |
Neurosphere-like human glioblastoma driven by CD133+ cancer stem cells
|
| Data processing |
Expression signals for each transcript and comparisons between different samples were calculated with the Affymetrix GeneChip® Operating Software GCOS1.4 and exported into Microsoft Excel.
|
| |
|
| Submission date |
Feb 27, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Christoph P. Beier |
| Organization name |
University of Regensburg
|
| Department |
Neurology
|
| Lab |
NeuroOncology
|
| Street address |
Universitatsstrasse 84
|
| City |
Regensburg |
| ZIP/Postal code |
93042 |
| Country |
Germany |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE7181 |
CD133+ and CD133- CSC lines in primary human GBM |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
