|
| Status |
Public on Apr 14, 2015 |
| Title |
B23 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Universal Reference provided by Miltenyi Biotec GmbH
|
| Organism |
synthetic construct |
| Characteristics |
sample type: syntethic miRNA pool of 954 miRNA sequences from human, mouse, rat, and virus
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol according to the manufacturer’s instructions.
|
| Label |
Hy3
|
| Label protocol |
2 µg of respective total RNA as well as 2 fmol of the Universal Reference 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 total RNA and Universal Reference mix was hybridized in a dual colour approach to microarrays.
|
| |
|
| Channel 2 |
| Source name |
B-D18
|
| Organism |
Homo sapiens |
| Characteristics |
sample type: hESC-H1-EB20-Day18 erythroid cells
cell type: erythroid cells
cell line: H1
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol according to the manufacturer’s instructions.
|
| Label |
Hy5
|
| Label protocol |
2 µg of respective total RNA as well as 2 fmol of the Universal Reference 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 total RNA and Universal Reference mix was 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 (TM) microarrays were done with the Agilent’s Microarray Scanner System (G2505C, Agilent Technologies, Palo Alto, USA) and ImaGene software Version 9.0 (BioDiscovery, Los Angeles, CA, USA).
|
| Data processing |
Signal quantification of the scanned miRXplore TM microarrays was done using ImaGene software Version 9.0 (BioDiscovery, Los Angeles, CA, USA). Local background was subtracted from the signal to obtain the net signal intensity and the ratio of Cy5/Cy3. 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. Within-array normalization occured by quantile normalization.
|
| |
|
| Submission date |
Mar 03, 2014 |
| Last update date |
Apr 14, 2015 |
| Contact name |
Hélène LAPILLONNE |
| E-mail(s) |
helene.lapillonne@trs.aphp.fr
|
| Organization name |
UPMC Univ Paris 06
|
| Department |
UMR_S938 CDR Saint-Antoine
|
| Lab |
Prolifération et Différentiation des Cellules Souches
|
| Street address |
27 Rue de Chaligny
|
| City |
Paris |
| ZIP/Postal code |
F-75012 |
| Country |
France |
| |
|
| Platform ID |
GPL15235 |
| Series (2) |
| GSE55530 |
Molecular Signature of Erythroblast Enucleation in Human Embryonic Stem Cells |
| GSE55532 |
Molecular Signature of Erythroblast Enucleation in Human Embryonic Stem Cells |
|
