|
| Status |
Public on Sep 02, 2011 |
| Title |
Bmi1 wild-type lung cells, biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Bmi1 wild-type lung cells
|
| Organism |
Mus musculus |
| Characteristics |
source tissue: lung
genetic background: Bl6/129 mixed background
genotype: Bmi1 wild-type
cell type: primary lung cells (CD31-negative, CD45-negative)
|
| Treatment protocol |
To make a total lung cell suspension, lungs were infused with dispase, then 1% agarose and lungs removed from animal; lungs were then minced, further digested with collagenase/dispase, treated with dnase, filtered, and red blood cells lysed. Cells were then labeled with 7aad, anti-CD31 and anti-CD45 antibodies, and FACS-purified to exclude endothelial, hematopoietic, and dead cells (See Kim et al, 2005).
|
| Growth protocol |
Primary lung cells were purified from 4 month old female mice; no ex vivo growth of cells prior to RNA extraction
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was purified using an Absolutely RNA Purification Kit (Agilent Technologies), and amplified using the WT-Ovation Pico RNA Amplification System (Nugen) to generate cDNA
|
| Label |
Biotin
|
| Label protocol |
The purified cDNA (5ug per sample) was fragmented and biotinylated using FL-Ovation cDNA Biotin Module V2 (Nugen, cat# 4200-12)
|
| |
|
| Hybridization protocol |
Fragmented, biotinylated cDNA was hybridized for 18 hours on Mouse 430_2 Affymetrix Mouse Genome 430 2.0 Arrays. The arrays were then washed and stained with streptavidin-phycoerythrin, using the Affymetrix GeneChip Fluidics Station 450
|
| Scan protocol |
Arrays were scanned using an Affymetrix GeneChip Scanner 3000
|
| Description |
Gene expression data from wild-type lung cells
|
| Data processing |
The raw intensity data were preprocessed by Affymetrix's Expression Console (version 1.1)
|
| |
|
| Submission date |
Jul 21, 2011 |
| Last update date |
Sep 02, 2011 |
| Contact name |
Carla Kim |
| E-mail(s) |
Carla.Kim@childrens.harvard.edu
|
| Organization name |
Children's Hospital Boston
|
| Department |
Hem/Onc, Stem Cell Program
|
| Lab |
Carla Kim Lab
|
| Street address |
300 Longwood Ave
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE30852 |
Expression data from Bmi1 mutant versus wild-type lung cells |
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