|
| Status |
Public on Mar 04, 2014 |
| Title |
CD34+ cells, SLPI shRNA 2 |
| Sample type |
RNA |
| |
|
| Source name |
CD34+ cells_SLPI shRNA
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: bone marrow of healthy individuals
cell type: CD34+ hematopoietic cells
transduced with: lenitvirus-based RFP-expressed shRNA constructs targeting SLPI mRNA
cell subtype: sorted RFP+ cells
|
| Treatment protocol |
After 3 days of the transductions, RFP+ cells were sorted, serum starved for 24 hours and treated with G-CSF 10 ng/ml for 18 hours. Total RNA of sorted cells was isolated and used for Affymetrix Exome arrays.
|
| Growth protocol |
Cells were cultured in Ex-vivo medium supplemented with 10 % HSA, 20 ng/ml IL-3, 20 ng/ml IL-6, 20 ng/ml TPO, 50 ng/ml SCF and 50 ng/ml Flt-3L.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using RNeasy Micro Kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
The Genechip WT cDNA Synthesis and Amplification Kit was used to make double-stranded cDNA from total RNA, which was then labeled with biotin (Genechip WT Terminal Labeling Kit).
|
| |
|
| Hybridization protocol |
After chemical fragmentation of the biotin-labeled cRNA targets, they were hybridized to Affymetrix Human Exome microarrays using the Fluidics Station 450
|
| Scan protocol |
Microarrays were scanned using the Affymetrix Genechip Scanner 3000 with Genechip Operating Software 1.4 (Affymetrix, Santa Clara, CA).
|
| Description |
SAMPLE 4
|
| Data processing |
CEL files were analyzed within Expression Console using gene-level RMA-Sketch default settings
|
| |
|
| Submission date |
Dec 16, 2013 |
| Last update date |
Mar 04, 2014 |
| Contact name |
Siarhei Kandabarau |
| E-mail(s) |
kandabarau.siarhei@mh-hannover.de
|
| Organization name |
Hannover Medical School
|
| Street address |
Carl-Neuberg-Str 1
|
| City |
Hannover |
| ZIP/Postal code |
30625 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE53353 |
A lack of secretory leukocyte protease inhibitor (SLPI) causes defects in granulocytic differentiation. |
|
