|
| Status |
Public on Aug 13, 2013 |
| Title |
(Biological replicate 3) E18.5 fetal cells sorted on CD31-CD45-Ter119-EpCAM++CD49f+ |
| Sample type |
RNA |
| |
|
| Source name |
E18.5 fetal cells sorted on CD31-CD45-Ter119-EpCAM++CD49f+
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl6J
gender: female
developmental stage: Embryonic day 18.5
tissue: mammary gland
cell type: E18.5 fetal cells sorted on CD31-CD45-Ter119-EpCAM++CD49f+
|
| Growth protocol |
CD31-CD45-Ter119-BP-1-EpCAM+CD49f+ adult basal cells and CD31-CD45-Ter119-EpCAM++CD49f+ E18.5 fetal cellswere isolated by FACS
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using the Absolutely RNA Nanoprep kit (Agilent) following the manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Fluorescent cRNA (complimentary RNA) was generated from 25 - 200 ng of RNA using Agilent's Low Input Quick Amp Labeling Kit. Prior to labeling, the integrity of the total RNA is checked with the Agilent 2100 Bioanalyzer (RNA), quantified with the NanoDrop ND-1000 UV-VIS spectrophotometer to measure A260/280 and A260/230 ratios or RiboGreen assay.Total RNA is initially converted into cDNA using a T7 polymerase primed reverse transcriptase reaction to synthesize first and second strand of cDNA. This cDNA then served as a template for cRNA synthesis by T7 RNA polymerase in the presence of cyanine 3-labeled CTP (Cy3).
|
| |
|
| Hybridization protocol |
1.65ug of Cy3 labeled cRNA is hybridized to the microarray at 65°C for 17 hours
|
| Scan protocol |
Preliminary fluorescence data is extracted from each microarray by the Agilent DNA Microarray Scanner
|
| Description |
E18.5_3
Gene expression from freshly isolated sorted cells
|
| Data processing |
The resulting data are processed by the Agilent Feature Extraction software (v.11.0.1.1). A QC report is generated for each microarray with a series of control panels evaluates the quality of the hybridization.
Green processed signal was quantile normalized with Agilent GeneSpring 12.0.
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| |
|
| Submission date |
Apr 24, 2013 |
| Last update date |
Aug 13, 2013 |
| Contact name |
Maisam Makarem |
| E-mail(s) |
maisam.makarem@gmail.com
|
| Organization name |
BC Cancer Research Center
|
| Street address |
675 W 10 Ave
|
| City |
Vancouver |
| State/province |
BC |
| ZIP/Postal code |
V5Z 1L3 |
| Country |
Canada |
| |
|
| Platform ID |
GPL10787 |
| Series (1) |
| GSE46357 |
Developmental changes in the in vitro activated regenerative activity of primitive mammary epithelial cells |
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