|
| Status |
Public on Oct 06, 2011 |
| Title |
Cyto_vs_Mito_1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
HeLa cells cytosolic fraction
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: HeLa
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cytosolic fraction using Trizol LS and from the mitochondrial fraction using Trizol following manufacturer's instructions
|
| Label |
Hy3
|
| Label protocol |
2 µg of respective total RNA were each mixed with 2.5 fmol of each of 18 RNA oligonucleotides reverse complement to miRControl 3 probes and subsequently fluorescently labelled using a commercial kit (miRCuryTM LNA microRNA Array Power labeling kit, Exiqon) following the instructions of the manufacturer. The corresponding total RNA samples were hybridized in a dual colour approach to microarrays.
|
| |
|
| Channel 2 |
| Source name |
HeLa cells mitochondrial fraction
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: HeLa
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cytosolic fraction using Trizol LS and from the mitochondrial fraction using Trizol following manufacturer's instructions
|
| Label |
Hy5
|
| Label protocol |
2 µg of respective total RNA were each mixed with 2.5 fmol of each of 18 RNA oligonucleotides reverse complement to miRControl 3 probes and subsequently fluorescently labelled using a commercial kit (miRCuryTM LNA microRNA Array Power labeling kit, Exiqon) following the instructions of the manufacturer. The corresponding total RNA samples were hybridized in a dual colour approach to microarrays.
|
| |
|
| |
| Hybridization protocol |
The fluorescently labeled samples were hybridized overnight to miRXplore TM Microarrays using the a-HybTM Hybridization Station (Miltenyi Biotec).
|
| Scan protocol |
Image capture and signal quantification of hybridized miRXplore ™ microarrays was done with the Agilent’s Microarray Scanner System (Agilent Technologies, Palo Alto, USA) and ImaGene software Version 8.0 (BioDiscovery, Los Angeles, CA, USA).
|
| Data processing |
Signal quantification of the scanned miRXplore TM microarrays was done using ImaGene software Version 8.0 (BioDiscovery, Los Angeles, CA, USA). Local background was subtracted from the signal to obtain the net signal intensity and the ratio of Hy5/Hy3. Subsequently, the mean of the ratios of 4 corresponding spots representing the same miRNA was computed. The mean ratios were normalised using the calibration oligos.
|
| |
|
| Submission date |
Oct 18, 2010 |
| Last update date |
Oct 06, 2011 |
| Contact name |
Silvia Rueberg |
| Organization name |
Miltenyi Biotec GmbH
|
| Department |
MACSmolecular Business Unit
|
| Lab |
Genomics Service Department
|
| Street address |
Friedrich-Ebert-Str. 68
|
| City |
Bergisch Gladbach |
| ZIP/Postal code |
51429 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10993 |
| Series (1) |
| GSE24761 |
HeLa cell mitochondrial microRNAs |
|
