|
| Status |
Public on Jan 01, 2014 |
| Title |
Prep1deltaSM_P58 |
| Sample type |
RNA |
| |
|
| Source name |
tibialis anterior muscle, Prep1deltaSM
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: C57BL/6
genotype/variation: Prep1deltaSM
gender: male
age: 8 weeks
tissue: tibialis anterior muscle
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using TRIzol reagent according to the manufacturer's instruction. RNA was subjected to DNase digestion to remove genomic DNA. Quantification of RNA was performed on a Nanodrop-1000 and RNA integrity was measured on Agilents Bioanalyzer.
|
| Label |
Cy3
|
| Label protocol |
Cy3-labeled cRNA was prepared from 1 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit from Agilent according to the manufacturer's instructions, followed by RNeasy column purification. Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturer's instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent-014868 Whole Mouse Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned using one color scan setting for 4x44k array slides.
|
| Description |
P58
RNA from tibialis muscle of Prep1deltaSM mice.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Nov 15, 2013 |
| Last update date |
Jan 01, 2014 |
| Contact name |
Timo Kanzleiter |
| Organization name |
DIfE
|
| Department |
Experimental Diabetology
|
| Street address |
Arthur-Scheunert-Alle 114
|
| City |
Nuthetal |
| ZIP/Postal code |
14558 |
| Country |
Germany |
| |
|
| Platform ID |
GPL7202 |
| Series (1) |
| GSE52424 |
Gene expression from Prep1-ablated mice |
|
