|
| Status |
Public on Dec 06, 2010 |
| Title |
Dyrk1a ES clone at T24, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
inducible ES cell clone
|
| Organism |
Mus musculus |
| Characteristics |
inducible gene: Dyrk1a
cell type: ES cell
|
| Treatment protocol |
Clones were grown in medium deprived of Tc to perform a time course of induction (24, 39 or 48hrs depending on experiment). In presence of Tc (0 hrs), the expression of each mRNA was indicative of the basal expression level in mES cells.
|
| Growth protocol |
ES cells were grown in mES media + LIF and supplemented with 1ug/ml tetracycline (Tc) at 37°C in an atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA samples at various times of induction were purified by QIAshredder and extracted with RNeasy Protect Mini Kit using standard conditions. Total RNA were reverse transcribed to single stranded cDNA with a special oligo (dT)24 primer containing a T7 RNA promoter site prior to second strand synthesis (One cycle cDNA synthesis kit by Affymetrix).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared using the GeneChip IVT labelling kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of biotinylated cRNA were hybridized for 16 hr at 45°C on GeneChip Mouse 430_2 Array (430A_2 in case of Runx1).
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Gene expression data from ES clone 24h after induction of gene overexpression.
|
| Data processing |
The data were analyzed using the RMA function of the Affy package of the Bioconductor tool. RMA analysis was performed separately on each set of six arrays relative to a same gene overexpression experiment.
|
| |
|
| Submission date |
Jan 11, 2010 |
| Last update date |
Dec 06, 2010 |
| Contact name |
Mario Lauria |
| E-mail(s) |
lauria@tigem.it
|
| Phone |
+39 081 6132 339
|
| Organization name |
TIGEM
|
| Street address |
via P. Castellino 111
|
| City |
Napoli |
| ZIP/Postal code |
80131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE19836 |
A mouse Embryonic Stem Cell Bank for inducible overexpression of human chromosome 21 genes |
|
