|
| Status |
Public on Jan 01, 2014 |
| Title |
Soleus Muscle_mKO_ZT0_replicate B |
| Sample type |
RNA |
| |
|
| Source name |
mKO_Soleus_ZT0
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6
genotype/variation: muscle-specific Bmal1 knockout (mKO)
age: male
gender: 5 months
tissue: soleus skeletal muscle
time point: Zeitgeber Time 0
source animal id: BM3 13
|
| Treatment protocol |
Animals were housed in an isolated temperature controlled room (22°C) under a 12-h light-dark regimen, with lights on at ZT0 (Zeitgeber Time 0, or 6am), lights off at ZT12 (Zeitgeber Time 12, or 6pm), with standard chow and water provided ad libitum. Soleus (SOL) and Tibialis Anterior (TA) were collected immediately after cervical dislocation at ZT0, 4, 8, 12, 16, and 20, snap frozen in liquid nitrogen and stored at -80°C until subsequent use.
|
| Growth protocol |
Animals: Conditional muscle-specific inactivation of Bmal1 (mKO) was obtained by crossing a mouse line with floxed Bmal1 (Storch et al., Cell, 2007) with mouse line carrying a Cre recombinase transgene under control of the Mlc1f promoter (Mlc1f-Cre) (Bothe et al., Genesis, 2000). 5 month old adult male mKO mice and Cre negative littermates (Ctrl) were used for microarray experiments.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
5.5μg sense strand DNA was prepared from 250ng total RNA using the Ambion® WT Expression Kit for Affymetrix® GeneChip® Whole Transcript (WT) Expression Arrays, and was fragmented and biotin-labeled using the GeneChip® WT Terminal Labeling Kit according to manufacturer's intructions.
|
| |
|
| Hybridization protocol |
Fragmented and biotin-labeled sense strand DNA were hybridized for 16 hr at 45C on Affymetrix Mouse Gene 1.0 ST arrays. Arrays were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
Standard Affymetrix scanning procedures on a GCS3000 7G Affymetrix scanner operated by GCOS, GeneChip Operating Software
|
| Description |
SOL_ZT0_KO_B
|
| Data processing |
RMA of .CEL files using affy Bioconductor library. In RMA, PM values have been background adjusted, normalized using quantile normalization, and expression measure calculated using median polish summarization
|
| |
|
| Submission date |
Dec 20, 2012 |
| Last update date |
Jan 01, 2014 |
| Contact name |
Silvio Bicciato |
| E-mail(s) |
[email protected]
|
| Phone |
+39-049-827-6108
|
| Organization name |
University of Padova
|
| Department |
Molecular Medicine
|
| Street address |
via U. Bassi 59/b
|
| City |
Padova |
| ZIP/Postal code |
35131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL13730 |
| Series (1) |
| GSE43071 |
Circadian transcriptome of fast tibialis anterior (TA) and slow soleus (SOL) skeletal muscles from muscle-specific Bmal1 KO and control Cre- littermates |
|
