|
| Status |
Public on Nov 24, 2010 |
| Title |
Macrophages_DD second day_CT4 |
| Sample type |
RNA |
| |
|
| Source name |
peritoneal macrophages, second day after release in DD, CT4
|
| Organism |
Mus musculus |
| Characteristics |
genetic background: C57BL/6
cell type: peritoneal macrophage
gender: male
time: second day CT4
|
| Growth protocol |
mice were entrained for two weeks in a 12 hour LD cycle before transferring them into DD. At indicated circadian times macrophages were harvested by lavage of the peritoneal cavity in 4hour intervalls and enriched by magnetic cell sorting based on the expression of the CD11b antigen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
total RNA was extracted by using the RNeasy Mini Kit (Qiagen). Total RNA of three mice per sampling time was pooled in equal amounts (48 ng/ µL).
|
| Label |
biotin
|
| Label protocol |
RNA biotin labeling was carried out using the Affimetrix GeneChip Whole Transcript Sense Target Labeling Assay according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
following cDNA synthesis, using random hexamers coupled with a T7 promotor sequence, obtained cRNA is converted into single stranded DNA, which then is fragmented by a treatment with a combination of uracil DNA glycosylase and apurinic/apyrimidinic endonuclease 1. Fragmented DNA was labeled by deoxynucleotidyl transferase reaction with labeling reagent that is linked to biotin. Hybridization and washing steps were done using the Affimetrix Hybridization and Wash and Stain Kit (according to manufacturer's instructions).
|
| Scan protocol |
Gene chips were scanned using the scanner type M10 (Pixel size 0.7, Filter 570).
|
| Data processing |
Data were analyzed by using the RMA gene default signal.
|
| |
|
| Submission date |
Nov 23, 2010 |
| Last update date |
Nov 23, 2010 |
| Contact name |
Bert Maier |
| E-mail(s) |
[email protected]
|
| Organization name |
Charité Universitätsmedizin
|
| Department |
Institute for Medical Immunology
|
| Lab |
Laboratory of Chronobiology
|
| Street address |
Hessische Straße 3-4
|
| City |
Berlin |
| ZIP/Postal code |
10115 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE25585 |
Circadian transcriptional profiling of peritoneal macrophages |
|
