|
| Status |
Public on Jul 14, 2010 |
| Title |
pCDNA/P19_aggregated |
| Sample type |
RNA |
| |
|
| Source name |
pCDNA/P19, aggregated state
|
| Organism |
Mus musculus |
| Characteristics |
cell type: embryonic carcinoma cell line
cell line: P19
genotype/variation: carrying an empty vector
|
| Treatment protocol |
To induce neural differentiation of these P19 cell lines, cells were allowed to aggregate in culture medium with 5×10-7 M all-trans-retinoic acid (RA) for 4 days. After aggregation, cells were replated onto cell culture grade dishes or poly-L-Lysine coated cell-disks for two days.
|
| Growth protocol |
P19 cells were cultured in alpha-modified Eagle’s essential medium (Sigma) containing 10% FCS and 2 mM L-glutamine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were prepared using ISOGENE (Nippon Gene, Toyama Japan) at each state of these cells according to the manufacture’s instruction. The qualities and integrities of these RNAs were verified by gel electrophoresis and by Agilent Bioanalyzer (Böblingen, Germany).
|
| Label |
Cy3
|
| Label protocol |
Cy3-labeled cRNA probes were synthesized according to the manufacture’s instruction (Agilent, Quick Amp Labeling Kit). Briefly mentioned, 0.2μg of each total RNA was reverse transcribed to double-stranded cDNA using T7 promoter sequence adding to oligo dT nucleotide as a primer. cRNAs were transcribed from these double-stranded cDNAs by T7 RNA polymerase.
|
| |
|
| Hybridization protocol |
Cy3-labeled cRNAs were hybridized to Whole Mouse Genome oligo-DNA microarray (Agilent) at 65°C for 17 hours using Gene Expression Hybridization Kit (Agilent) and washed using Gene Expression Wash Pack (Agilent).
|
| Scan protocol |
Intensities of Cy3-fluorescent signals were scanned by Agilent DNA microarray scanner.
|
| Description |
Gene expression of pCDNA/P19 cells in the aggregated state
|
| Data processing |
The raw data was converted into the quantitative data using Agilent Feature Extraction software version 10.1.1.1. Data were further normalized using GeneSpringGX10 using Median shift normalization to the 75th percentile and baseline transformed to the median of all samples.
|
| |
|
| Submission date |
Jul 02, 2010 |
| Last update date |
Jul 13, 2010 |
| Contact name |
Kohzo Nakayama |
| E-mail(s) |
kohzona@ shinshu-u.ac.jp
|
| Phone |
+81-263-37-2594
|
| Fax |
+81-263-37-2594
|
| Organization name |
Shinshu University
|
| Department |
Anatomy
|
| Street address |
Asahi
|
| City |
Matsumoto |
| ZIP/Postal code |
390-8621 |
| Country |
Japan |
| |
|
| Platform ID |
GPL7202 |
| Series (1) |
| GSE22690 |
The intracellular domain of amyloid precursor protein induces the dynamic change of the gene expression profile in P19 cells. |
|
