|
| Status |
Public on Jun 05, 2018 |
| Title |
ivHSC_biological rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Fresh umbilical cord blood
|
| Organism |
Homo sapiens |
| Characteristics |
blood fraction/population: fHSC population
cell type: ivHSC
|
| Treatment protocol |
Cells were labeled using CD34, CD38, CD71 and CD45RA by purification using FACS. Recovery of cells was made using PBS 1X + EDTA 2mM + 2% BFS. Before RNA extraction, cells were washed using PBS and placed in lysis solution quickly.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was made according to the protocol for Single cell RNA purification Kit by Norgen Biotek Cat. 51800.
|
| Label |
biotin
|
| Label protocol |
The WT Pico Reagent Kit was used to generate the biotinylated cRNA starting from 100pg of total RNA according to the manufacturer (Manual Target preparation for GeneChip Whole transcript (WT) expression arrays, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 20 µg cRNA was hybridized for 16 hrs at 45ºC to 60 RPM on GeneChip Human Transcriptome Array V2. GeneChip was washed and stained in the station Fluidics Station 450 (Affymetrix, Santa Clara, USA).
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 7G (Affymetrix , Santa Clara, USA).
|
| Description |
ivHSC2
Gene expression data from in vitro generated HSC from fresh HSC took from USB.
|
| Data processing |
Array signal intensities were analyzed with the Affymetrix Expression Console using Signal Space Transformation-Robust Multichip Analysis (SST-RMA) for the background correction.
|
| |
|
| Submission date |
Nov 29, 2017 |
| Last update date |
Jun 05, 2018 |
| Contact name |
Hector Mayani |
| E-mail(s) |
[email protected]
|
| Phone |
+5215543984800
|
| Organization name |
Oncology Research Unit
|
| Lab |
Hematopoietic Stem Cells Laboratory
|
| Street address |
Cuahutemoc 330
|
| City |
Mexico |
| State/province |
Mexico City |
| ZIP/Postal code |
06720 |
| Country |
Mexico |
| |
|
| Platform ID |
GPL17586 |
| Series (1) |
| GSE107497 |
Genomic analysis for hematopoietic stem and progenitors cells (HSPC) generated in vitro according to ex vivo expansion protocols and their comparison with HSPC obtained fresh |
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