|
| Status |
Public on Dec 23, 2018 |
| Title |
RL-/- low rep4 |
| Sample type |
RNA |
| |
|
| Source name |
bone marrow
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6
age: 8-10 weeks
tissue: monocytes
genotype: RL-/-
ly6c_expression: low
|
| Treatment protocol |
After Fc receptor block (CD16/32), cells were labelled with CD3e-PE, CD11b-Alexa488, CD19-PE, CD49b-PE, CD117-PE, Ly6C-PerCP-Cy5.5, Ly6G-PE and NK1.1-PE for 30 min on ice in the dark. DAPI was added shortly before measurements. Monocyte subsets were sorted by a MoFlo Legacy cell sorter from Beckman Coulter.
|
| Growth protocol |
For the isolation of monocytes, femurs and tibiae were taken, bone marrow was flushed out with RPMI 1640 +1% FCS, red blood cell lysis was performed and cells were filtered through a 70 µm mesh to obtain a single cell suspension.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using RNeasy mini kit from Qiagen, RNA concentration was increased by evaporation for 18 min in a SpeedVac system. RNA integrity was analyzed using the Agilent RNA 6000 Nano total RNA kit and an Agilent 2000 Bioanalyzer system. RNA was transcribed into cDNA with SuperScript III Reverse Transcriptase (ThermoFisher Scientific).
|
| Label |
biotin
|
| Label protocol |
Biotin-aRNA was made using the TargetAmp-Nano Labeling kit for Illumina Expression BeadChip from Epicentre.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol.
|
| Scan protocol |
Standard Illumina scanning protocol using an Illumina BeadArray Reader.
|
| Description |
replicate 4
252
|
| Data processing |
The data was normalised using quantile normalisation with Illumina GenomeStudio.
|
| |
|
| Submission date |
Sep 11, 2017 |
| Last update date |
Dec 23, 2018 |
| Contact name |
Frank Rühle |
| E-mail(s) |
f.ruehle@imb-mainz.de
|
| Organization name |
Institute of Molecular Biology (IMB)
|
| Department |
Bioinformatics Core Facility
|
| Street address |
Ackermannweg 4
|
| City |
Mainz |
| ZIP/Postal code |
55128 |
| Country |
Germany |
| |
|
| Platform ID |
GPL18752 |
| Series (1) |
| GSE103705 |
Suppressor of cytokine signaling 1 is involved in gene regulation which control the survival of Ly6Clow monocytes in mice |
|
