|
| Status |
Public on Dec 31, 2009 |
| Title |
NS5_1 |
| Sample type |
RNA |
| |
|
| Source name |
NS5 Neural Stem Cells
|
| Organism |
Mus musculus |
| Characteristics |
cell line
|
| Treatment protocol |
For differentiation into GFAP-positive cells, NS5s were seeded at 5x104 cells/cm2 in normal growth medium (NGM) on gelatin-coated plastic or glass coverslips overnight before medium was switched into NGM in the absence of growth factors and with the addition of 5% FCS for 3 days. Cells were routinely passaged with a 1:5-10 split using trypsin/EDTA
|
| Growth protocol |
NS5 cells were grown as described in Conti et al, PLoS Biology Vol. 3, No. 9, e283 doi:10.1371/journal.pbio.0030283
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were detached from tissue culture plates, pelleted and lysed with TRIzol reagent (Invitrogen, Cat# 15596-018) as per kit instruction. The resultant total RNA extracts were further cleaned up with Qiagen RNEasy Mini Kit (Qiagen Cat# 74106) as per kit instruction
|
| Label |
biotin
|
| Label protocol |
4 biological replicates of total RNA extracted from each of NS5 and Astrocyte cells were subjected to reverse transcription, second-strand cDNA synthesis and in vitro transcription using Illumina TotalPrep RNA Amplification Kit (Ambion, USA, Cat# IL1791). Biotin was incorporated into amplified RNA through use of biotin-modified dUTP.
|
| |
|
| Hybridization protocol |
Amplified RNA were hypridised on Sentrix Mouse Ref-8 (v1.1) Expression BeadChips (Illumina, USA, Cat# 11234312) according to Illumina protocols
|
| Scan protocol |
Chips were washed and scanned according to Illumina protocols
|
| Description |
NS5_1
|
| Data processing |
Quantile normalisation, log2 transformation using the beadarray Bioconductor library in R
|
| |
|
| Submission date |
Mar 04, 2008 |
| Last update date |
May 28, 2009 |
| Contact name |
Caroline Emma Johnston |
| E-mail(s) |
caroline.johnston@iop.kcl.ac.uk
|
| Organization name |
Kings College London
|
| Street address |
125 Coldharbour Lane
|
| City |
London |
| ZIP/Postal code |
SE5 9NU |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6103 |
| Series (1) |
| GSE10721 |
Differential gene expression in murine neural stem cells (NS5) and neural stem cell-derived astrocytes |
|
