|
Status |
Public on Jan 23, 2010 |
Title |
fNSC rep1 |
Sample type |
RNA |
|
|
Source name |
female NSC
|
Organism |
Mus musculus |
Characteristics |
gender: female cell type: neural stem cell
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA samples to be analyzed on microarrays were prepared using Qiagen RNeasy columns with on-column DNA digestion.
|
Label |
biotin
|
Label protocol |
Specifically, 400 ng of total RNA per sample was used as input into a linear amplification protocol for labelled cRNA synthesis (Illumina TotalPrep RNA Amplification Kit - Ambion) following the manufacturer's instructions, which involved synthesis of T7-linked double-stranded cDNA and 10-12 hrs of in-vitro transcription (IVT) incorporating biotin-labelled nucleotides.
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|
|
Hybridization protocol |
Standard Illumina hybridization protocol. Purified and labelled cRNA was then hybridized for 18 hrs onto MouseRef-8 v2 expression BeadChips (Illumina) according to the manufacturer's instructions. After washing, as recommended, chips were stained with streptavidin-Cy3 (GE Healthcare). Samples were exclusively hybridized as biological replicates.
|
Scan protocol |
Standard Illumina scanning protocol. Scanned using the iScan reader (Illumina) and accompanying software.
|
Description |
fNSC: Mouse female NSC (Neural Stem Cell). Replicate 1.
|
Data processing |
The bead intensities were mapped to gene information using BeadStudio 3.2 (Illumina), background correction was perform using the Affymetrix Robust Multi-array Analysis (RMA) background correction model, variance stabilization was performed using the log2 scaling and gene expression normalization was calculated with the quantile method implemented in the lumi package of R-Bioconductor.
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|
|
Submission date |
Apr 06, 2009 |
Last update date |
Jan 15, 2022 |
Contact name |
Marcos J. Araúzo-Bravo |
E-mail(s) |
[email protected]
|
Phone |
+34 943 00 6108
|
Organization name |
Max Planck Institute for Molecular Biomedicine
|
Department |
Cell and Developmental Biology
|
Lab |
Computational Biology and Bionformatics
|
Street address |
Rogentstrasse
|
City |
Muenster |
ZIP/Postal code |
48149 |
Country |
Germany |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE15561 |
Generation of parthenogenetic iPS cells from parthenogenetic neural stem cell |
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