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Status |
Public on Dec 14, 2012 |
Title |
EBTPd12#2 |
Sample type |
RNA |
|
|
Source name |
CD34+CD43+CD45+ cells/EBd12
|
Organism |
Homo sapiens |
Characteristics |
cell type: CD34+CD43+CD45+ hematopoietic stem/progenitor cells days of eb differentiation: 12
|
Growth protocol |
ES cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM)-F12 (Sigma) supplemented with 1x MEM non-essential amino acids (Gibco-Invitrogen), 1x GlutaMAX-I (Gibco-Invitrogen), 20% knockout serum replacement (KSR) (Gibco-Invitrogen), 0.1 mM 2-mercaptoethanol (2-ME)(Sigma), and 5 ng/mL basic fibroblast growth factor (bFGF)(ReproCELL) either on irradiated MEF as feeder layers.
|
Extracted molecule |
total RNA |
Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
Label |
biotin
|
Label protocol |
45 μL of fragmented single-stranded DNA was mixed with 15 μL of master mix containing 5X TdT buffer, TdT and DNA Labeling reagent, and was incubated at 37°C for 60 minutes.
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|
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Hybridization protocol |
The hybridization cocktail was denatured at 99°C for 5 minutes, incubated at 45°C for 5 minutes, and then injected into a GeneChip ST cartridge. The arrays were incubated at 45°C for 17 hours ± 1 hour in a rotating oven at 60 rpm. The arrays were then washed and stained according to the GeneChip Expression Wash, Stain and Scan User Manual (P/N702731).
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Scan protocol |
Fluorescence signals were scanned with the Affymetrix GeneChip Scaner 3000. Affymetrix GeneChip Command Console (AGCC) software was used for acquisition of the initial image of the scanned array.
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Description |
CD34+CD43+CD45+ cells differentiated for 12 days in EBs
|
Data processing |
The Affymetrix Expression Console was used to normalize data and determine signal intensity (RMA-Sketch).
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|
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Submission date |
Apr 17, 2012 |
Last update date |
Dec 14, 2012 |
Contact name |
Takaho A. Endo |
E-mail(s) |
[email protected]
|
Organization name |
RIKEN
|
Department |
IMS
|
Lab |
Laboratory for Integrative Genomics
|
Street address |
1-7-22 Suehiro, Tsurumi
|
City |
Yokohama |
State/province |
Kanagawa |
ZIP/Postal code |
230-0045 |
Country |
Japan |
|
|
Platform ID |
GPL6244 |
Series (2) |
GSE37348 |
Expression data of human ES cells-derived CD34+CD43+CD45low cells (hemogenic endothelium-like cells) expanded upon overexpression of Sox17 |
GSE38156 |
Critical role of SOX17 in the hematopoietic development from human embryonic stem cells |
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