|
| Status |
Public on May 26, 2011 |
| Title |
CDM 1 |
| Sample type |
RNA |
| |
|
| Source name |
muscle primary culture cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: myotube
disease state: congenital myotonic dystrophy
|
| Treatment protocol |
Biopsies from quadriceps muscles were obtained during autopsies of control or age-matched CDM1 patient.
|
| Growth protocol |
Myoblasts were grown in HAM’s F10 medium (Gibco) supplemented with 50 µg/ml gentamycine (Biomedia) and 20% fetal calf serum (Biomedia). All cultures were incubated at 37°C in a humid atmosphere containing 5% CO2. For myoblast differentiation, growth medium was removed from sub-confluent cultures and replaced by Dulbecco’s modified Eagle’s medium (Gibco) containing 10 µg/ml insulin and 100 µg/ml transferrin (Sigma)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
TRIZOL
|
| Label |
Cy3
|
| Label protocol |
Each sample was prepared using Agilent’s “miRNA Complete Labeling and Hyb” Kit (5190-0456) according to the Manufacturer’s protocol. Total RNA (100 ng) was dephosphorylated with Calf Intestinal Alkaline Phosphatase at 37°C for 30 min, denatured with DMSO at 100°C for 10 min, and labelled with Cyanine3-pCp using T4 RNA ligase at 16°C for 2 h.
|
| |
|
| Hybridization protocol |
The labeled RNAs were hybridized to Human miRNA V2 8x15K microarrays (G4470B, Agilent) at 55°C for 20 h in a Agilent rotating oven.
|
| Scan protocol |
After washing, the arrays were scanned with an Agilent Microarray Scanner (G2565B) using extended dynamic range as specified by the manufacturer.
|
| Data processing |
Agilent Feature Extraction 9.5 Software was used to extract the microarray data using default parameters (protocol miRNA-v1_95_May07 and Grid: 019118_D_20080214)
|
| |
|
| Submission date |
Sep 13, 2010 |
| Last update date |
May 27, 2011 |
| Contact name |
Doulaye Dembele |
| E-mail(s) |
doulaye@igbmc.fr
|
| Phone |
+33 3 88 65 35 28
|
| Organization name |
IGBMC
|
| Department |
Biopuces
|
| Street address |
1 rue Laurent Fries
|
| City |
Illkirch |
| ZIP/Postal code |
67400 |
| Country |
France |
| |
|
| Platform ID |
GPL7731 |
| Series (1) |
| GSE24109 |
Antagonistic role of MBNL1 and LIN28 promotes specific alteration of pre-miR-1 processing and is associated with heart defects in DM |
|
