|
| Status |
Public on Jan 01, 2020 |
| Title |
Control_1 |
| Sample type |
RNA |
| |
|
| Source name |
CD34+ HSPC ctrl day 5 #1
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD34+ HSPC in monocytic differentiation medium
lentiviral construct: empty control vector
|
| Treatment protocol |
NA
|
| Growth protocol |
Three independent samples of human CD34+ HSPC from cord blood were transduced with lentiviral constructs containng HOXA10-AS or with a empty control vector. Transduced cells were FACS sorted for EGFP expression and cultured in conditions promoting myeloid differentiation. Total RNA was prepared of FACS-sorted CD11b+CD14-CD117- myeloid precursors on 5 of culture. We analyzed early gene expression changes using Affymetrix Microarray Clariom™ S Pico Assay.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Zymo Research Quick RNA Miniprep Kit according to manufacturers instructions. RNA quality was assessed using the Agilent 2100 Bioanalyzer.
|
| Label |
Biotin
|
| Label protocol |
2-10 ng of total RNA were used for biotin labelling according GeneChip® Pico Kit (Affymetrix). 5.5 μg of biotinylated cDNA were fragmented and placed in a hybridization cocktail containing four biotinylated hybridization controls (BioB, BioC, BioD, and Cre) as recommended by the manufacturer. Samples were hybridized to an identical lot of Affymetrix Clariom™ S (400 Format) for 17 hours at 45°C.
|
| |
|
| Hybridization protocol |
Hybridization was done for 16 hrs at conditions recommended by the manufacturer. Clariom™ S chips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GCS 3000.
|
| Description |
Gene expression data from CD11b+CD117-CD14- myeloid precursors transduced with ctrl vector
|
| Data processing |
Image analysis was done by Affymetrix® GeneChip® Command Console® Software (AGCC) and Affymetrix® Expression Console™ Software.
|
| |
|
| Submission date |
Dec 12, 2018 |
| Last update date |
Jan 02, 2020 |
| Contact name |
Jan-Henning Klusmann |
| E-mail(s) |
jan-henning.klusmann@kgu.de
|
| Organization name |
Goehte University Frankfurt
|
| Department |
Pediatric Hematology and Oncology
|
| Street address |
Theodor-Stern-Kai 7
|
| City |
Frankfurt |
| ZIP/Postal code |
60590 |
| Country |
Germany |
| |
|
| Platform ID |
GPL23159 |
| Series (1) |
| GSE123694 |
Gene expression data of human chord blood hematopoietic progenior cells subjected to myeloid differentiation and transduced with lncRNA HOXA10-AS |
|
