|
| Status |
Public on Apr 11, 2018 |
| Title |
corpuscallosum_untreated_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
Corpus callosum, untreated controls, replicate 4
|
| Organism |
Mus musculus |
| Characteristics |
tissue: corpus callosum
gender: male
treatment: untreated
|
| Treatment protocol |
To induce demyelination, cuprizone (CPZ) was administrated for four weeks orally. The powdered standard chow was mixed with 0.2-0.4% CPZ. After 4 weeks, treatment with cuprizone diet was stopped. Remyelination was examined at two time points: acute remyelination induced by four weeks feeding with CPZ followed by two days of regular diet and full remyelination induced by four weeks CPZ feeding followed by two weeks of regular diet. Control mice were kept on a normal diet.
|
| Growth protocol |
0
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The affected areas of corpus callosum was punched out and RNA was extracted by the miRNeasy micro Kit (Qiagen, Valencia, CA). The quantity and quality of total RNA was assessed by NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE) and Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA), respectively. Only those samples that gave >8.0 for RNA integrity number, showed a clear gel image and no DNA contamination was observed on the histogram were used for microarray experiments.
|
| Label |
Cy3
|
| Label protocol |
100 ng of total RNA was first dephosphorylated by using calf intestine alkaline phosphatase and labeled by using miRNA labeling Reagent & Hybridization Kit according to the manufacturer’s protocol with no modification.
|
| |
|
| Hybridization protocol |
The labeled samples were hybridized for 20 hours at 55°C to Agilent Mouse miRNA Microarrays (G4472A, 8x15k) in a rotating Agilent hybridization oven.
|
| Scan protocol |
The arrays were scanned with an Agilent DNA Microarray Scanner BA.
|
| Description |
miRNA expression
|
| Data processing |
Signal quantification was carried out by Feature Extraction (ver. 10.5.1.1) using default parameters (protocol: miRNA_105_Dec08 and Grid: 019119_D_F_20081129). Image Analysis Software and data were further analyzed by Genespring GX10.0.
|
| |
|
| Submission date |
Jun 29, 2017 |
| Last update date |
Apr 11, 2018 |
| Contact name |
Viktor Molnár |
| E-mail(s) |
molvik.dgci@gmail.com
|
| Organization name |
Semmelweis University
|
| Department |
Department of Genetics, Cell- and Immunobiology
|
| Street address |
Nagyvárad tér 4.
|
| City |
Budapest |
| ZIP/Postal code |
1089 |
| Country |
Hungary |
| |
|
| Platform ID |
GPL8824 |
| Series (1) |
| GSE100662 |
MicroRNA profiling of demyelination and remyelination areas in corpus callosum of cuprizone-treated mice |
|
