|
| Status |
Public on Dec 24, 2013 |
| Title |
Bone marrow macrophages 3 days CSF-1 versus 0 days ERT2 Cre Thoc5 (flox/flox) with tamoxifen rep1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Bone marrow derived macrophages harvested at day 0 of differentiation induction
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Bone marrow macrophages
genotype: ERT2 Cre Thoc5 (flox/flox)
differentiation stage: day 0
treatment: none
genetic background: C57BL/6 x B6:129 Ola
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cytoplasmic RNA was purified by use of the Pure RNA isolation Kit (Roth).
|
| Label |
Alexa fluor 555
|
| Label protocol |
Synthesis of Cy3- or Cy5-labeled cRNA was performed with the “Amino Allyl MessageAmp™ II aRNA Amplification Kit” (#5190-0444, Life Technologies) according to the manufacturer’s recommendations, except that the molar proportion of aminoallyl-UTP and UTP used was adjusted to 1+11 (instead of 1+1).
|
| |
|
| Channel 2 |
| Source name |
Bone marrow derived macrophages harvested at day 3 after differentiation induction
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Bone marrow macrophages
genotype: ERT2 Cre Thoc5 (flox/flox)
differentiation stage: day 3 (under CSF-1)
treatment: tamoxifen
genetic background: C57BL/6 x B6:129 Ola
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cytoplasmic RNA was purified by use of the Pure RNA isolation Kit (Roth).
|
| Label |
Alexa fluor 647
|
| Label protocol |
Synthesis of Cy3- or Cy5-labeled cRNA was performed with the “Amino Allyl MessageAmp™ II aRNA Amplification Kit” (#5190-0444, Life Technologies) according to the manufacturer’s recommendations, except that the molar proportion of aminoallyl-UTP and UTP used was adjusted to 1+11 (instead of 1+1).
|
| |
|
| |
| Hybridization protocol |
cRNA fragmentation, hybridization and washing steps were carried-out exactly as recommended in the “Two-Color Microarray-Based Gene Expression Analysis Protocol V5.7” (Agilent Technologies).
|
| Scan protocol |
Slides were scanned on the Agilent Micro Array Scanner G2565CA (pixel resolution 5 µm, bit depth 20).
|
| Data processing |
Data extraction, processing and intra-array normalization of raw fluorescence intensity values were performed with the “Feature Extraction Software V10.7.3.1” by using the recommended default extraction protocol file: GE2_107_Sep09.xml.
|
| |
|
| Submission date |
Sep 26, 2012 |
| Last update date |
Dec 24, 2013 |
| Contact name |
Oliver Dittrich-Breiholz |
| E-mail(s) |
dittrich.oliver@mh-hannover.de
|
| Organization name |
Medical School Hannover
|
| Department |
Research Core Unit Genomics
|
| Street address |
Carl-Neuberg-Str. 1
|
| City |
Hannover |
| ZIP/Postal code |
30625 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10333 |
| Series (1) |
| GSE41170 |
THOC5, a member of the mRNA export complex, is essential for hematopoiesis in vivo and is required for CSF-1 induced macrophage differentiation |
|
