|
Status |
Public on Oct 30, 2010 |
Title |
Erythroid |
Sample type |
SRA |
|
|
Source name |
Primary pro-erythroblasts derived ex vivo from CD34+ multipotential hematopoietic progenitors
|
Organism |
Homo sapiens |
Characteristics |
cell type: Pro-erythroblasts chip antibody: anti-TAL1 Ab (Santa Cruz sc-12984)
|
Treatment protocol |
Human primary proerythroblasts were obtained by ex vivo differentiation from CD34+ hematopoietic stem cells. Briefly, light-density mononuclear cells were isolated by ficoll density gradient centrifugation from G-CSF-mobilized adult blood from donors without hematological malignancies (Ottawa Hospital Research Ethics Board #2007804-01H). CD34+ cells were enriched through positive immunomagnetic selection using the CD34 MicroBead Kit (Miltenyi Biotec Inc.) (purity > 95 ± 3%) and differentiated ex vivo as described in (Giarratana et al., 2005) except that we used 20% BIT (StemCell Technologies). Cells at day 10 of differentiation (corresponding to the proerythroblast stage) were used for chromatin immunoprecipitation
|
Extracted molecule |
genomic DNA |
Extraction protocol |
ChIPed DNA was prepared according to the Illumina protocol with two modifications: 1) DNA fragments ranging from 150bp to 300bp were selected at the gel selection step; 2) 21 instead of 18 cycles of PCR were done at the amplification step.
|
|
|
Library strategy |
ChIP-Seq |
Library source |
genomic |
Library selection |
ChIP |
Instrument model |
Illumina Genome Analyzer |
|
|
Description |
Human primary proerythroblasts were obtained by ex vivo differentiation from CD34+ hematopoietic stem cells. Briefly, light-density mononuclear cells were isolated by ficoll density gradient centrifugation from G-CSF-mobilized adult blood from donors without hematological malignancies (Ottawa Hospital Research Ethics Board #2007804-01H). CD34+ cells were enriched through positive immunomagnetic selection using the CD34 MicroBead Kit (Miltenyi Biotec Inc.) (purity > 95 ± 3%) and differentiated ex vivo as described in (Giarratana et al., 2005) except that we used 20% BIT (StemCell Technologies). Cells at day 10 of differentiation (corresponding to the proerythroblast stage) were used for chromatin immunoprecipitation
|
Data processing |
Sequence reads were obtained using the Illumina Genome Analyzer II. Reads were aligned to human genome (hg18) using MAQ (version 0.6.6) .
|
|
|
Submission date |
Oct 28, 2010 |
Last update date |
May 15, 2019 |
Contact name |
Stephen Tapscott |
E-mail(s) |
stapscot@fredhutch.org
|
Organization name |
Fred Hutch Cancer Research Center
|
Department |
Human Biology
|
Lab |
Tapscott
|
Street address |
1100 Fairview N. Ave
|
City |
Seattle |
State/province |
WASHINGTON |
ZIP/Postal code |
98103 |
Country |
USA |
|
|
Platform ID |
GPL9052 |
Series (1) |
GSE25000 |
Differential genomic targeting of the transcription factor TAL1 in alternate hematopoietic lineages |
|
Relations |
SRA |
SRX029598 |
BioSample |
SAMN00116839 |