|
| Status |
Public on Aug 01, 2011 |
| Title |
CD70Tg biological rep 3 |
| Sample type |
RNA |
| |
|
| Source name |
CD71+ bone marrow cells from CD70TG (clone F13) mouse 3
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
genotype/variation: CD70TG
tissue: Bone marrow
cell type: CD71+ cells (MACS-enriched)
|
| Treatment protocol |
Single cell suspensions from bone marrow were obtained by crushing the bones of the hindlegs, and filtering the suspension through a 40µm cell strainer. CD71+ cells were MACS-enriched from the bone marrow of WT, CD70TG, IFNg-/- and CD70TG*IFNg-/- mice by labelling with biotin-conjugated CD71 antibodies (eBioscience) and using streptavidin-conjugated microbeads (Miltenyi Biotech).
|
| Growth protocol |
Mice were bred and maintained on a C57BL/6 background in the animal facilities of the Academic Medical Center (University of Amsterdam, Amsterdam, The Netherlands) in specific pathogen-free conditions. Mice were given standard chow and acidified drinking water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
A comprehensive description of step-by-step instructions and lists of reagents required for the optimized Whole Transcript (WT) Sense Target Labeling Assay are described in the GeneChip Whole Transcript Sense Target Labeling Assay Manual (P/N 701880), available at www.affymetrix.com.
|
| |
|
| Hybridization protocol |
FS450_0001
100-300ng of total RNA was hybridized to Mouse Gene 1.0 ST Array GeneChips, according to the manufacturer’s protocols (Affymetrix).
|
| Scan protocol |
According to affymetrix scan protocol, available at www.affymetrix.com using a Scanner 3000 7G. P/N 702569 Rev. 5
|
| Description |
Gene expression in CD71+ bone marrow cells from CD70TG mice
|
| Data processing |
RMA pre-processing and baseline transformation to median of all samples using Genespring GX Software (Agilent Technologies).
|
| |
|
| Submission date |
Jul 01, 2010 |
| Last update date |
Aug 01, 2011 |
| Contact name |
Laura Gutierrez |
| E-mail(s) |
l.gutierrez@sanquin.nl
|
| Organization name |
Sanquin Research
|
| Department |
Blood Cell Research
|
| Street address |
Plesmanlaan 125
|
| City |
Amsterdam |
| ZIP/Postal code |
1066CX |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE22656 |
Expression data from CD71+ cells from the bone marrow of WT, CD70TG, IFNg-/- and CD70TG*IFNg-/- mice. |
|
