|
| Status |
Public on Apr 05, 2011 |
| Title |
LSK EZH2 REP3 |
| Sample type |
RNA |
| |
|
| Source name |
LSK EZH2
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6J
cell type: LSK
genotype/variation: EZH2 overexpression
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA isolated with Trizol.
Total RNA fraction. RNA Integrity Numbers were in the range 8.6 to 9.4 when assayed by Lab-chip technology on an Agilent 2100 Bioanalyzer.
|
| Label |
Cy3
|
| Label protocol |
Amount of nucleic acid labeled: 30ng. Amplification: by RNA polymerases. Commercial One-Color Microarray-Based Gene Expression Analysis (Low Input Quick Amp Labeling) kit by following manufacturer instructions. Agilent manual G4140-90040 Ver 6.5 May 201
Briefly, MMLV-RT retrotranscription of sample from a T7 promoter primer is followed by a T7 RNA pol catalysed in vitro transcription reaction in the presence of Cy3-CTP fluorophore.
Labeled samples are purified with silica-based RNeasy spin columns (Qiagen).
|
| |
|
| Hybridization protocol |
Chamber type: SureHyb hybridization chamber (Agilent). Quantity of each labeled extract used: 600 ng. Volume: 40 uL. Temperature (ºC): 65. Duration: 17 hours.
|
| Scan protocol |
Scanned on an G2565C DNA microarray scanner (Agilent). Images were quantified using Agilent Feature Extraction Software (ver. 10.7).
|
| Description |
EZH2-cKI-3
|
| Data processing |
Normalized signal intensities were obtained by a) background subtraction, b) threshold raw signals to 1, c) log base 2 transformation, d) global intra-array normalization to the 75.0 percentile, and e) per-gene baseline transformation by subtracting the median for that same gene in all samples.
|
| |
|
| Submission date |
Apr 04, 2011 |
| Last update date |
Apr 06, 2011 |
| Contact name |
Antonio Herrera Merchan |
| E-mail(s) |
aherrera@cnic.es
|
| Organization name |
CNIC
|
| Department |
Regenerative Cardiology
|
| Lab |
Stem Cell Aging
|
| Street address |
Melchor Fernández Almagro, 3
|
| City |
Madrid |
| ZIP/Postal code |
28029 |
| Country |
Spain |
| |
|
| Platform ID |
GPL10787 |
| Series (1) |
| GSE28369 |
Expression data from LSK mice that overpexpress Ezh2 versus LSK Control |
|
