|
| Status |
Public on Aug 12, 2008 |
| Title |
7561MLB25LN | NoDox |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
murine lymphoma cell line, 7561MLB25LN, NoDox
|
| Organism |
Mus musculus |
| Characteristics |
murine lymphoma cell line | 7561MLB25LN
18 hours
Tissue: lymphocytes
Strain: NMRI mice
|
| Treatment protocol |
NoDox
|
| Growth protocol |
cell lines were cultured in RPMI medium 1640 with 10% FCS, 1% Penicillin/Streptomycin, 1% L-Glutamin, 1% non-essential amino acids, and 0.1% b-Mercaptoethanol
|
| Extracted molecule |
total RNA |
| Extraction protocol |
total RNA was isolated using the mirVana miRNA Isolation Kit (Ambion) followed by isolating size-fractionated small RNA using the flashPAGE Fractionator System (Ambion)
|
| Label |
Cy5
|
| Label protocol |
end-labeling strategy (mirVana miRNA Labeling Kit, Ambion)
|
| |
|
| Channel 2 |
| Source name |
pool of RNA derived from differentiated wild type mouse tissues
|
| Organism |
Mus musculus |
| Characteristics |
Strain: NMRI
18 hours
Tissue: brain/lung/thymus/spleen/kidney/heart/ovary/testis/liver
|
| Treatment protocol |
18 hours Doxycycline (Dox) treatment (1µg/ml)
|
| Growth protocol |
cell lines were cultured in RPMI medium 1640 with 10% FCS, 1% Penicillin/Streptomycin, 1% L-Glutamin, 1% non-essential amino acids, and 0.1% b-Mercaptoethanol
|
| Extracted molecule |
total RNA |
| Extraction protocol |
total RNA was isolated using the mirVana miRNA Isolation Kit (Ambion) followed by isolating size-fractionated small RNA using the flashPAGE Fractionator System (Ambion)
|
| Label |
Cy3
|
| Label protocol |
end-labeling strategy (mirVana miRNA Labeling Kit, Ambion)
|
| |
|
| |
| Hybridization protocol |
co-hybridization of lymphoma cell lines and common reference was performed for 14 hrs onto the miRNA microarrays and washed according to the manufacturer’s protocol (Ambion)
|
| Scan protocol |
Microarrays were imaged using an Axon GenePix 4000B laser scanner (Axon Instruments). Fluorescence ratios were extracted (tumor/common reference) after subtracting the background using the GenePix Pro 6.0 software (Axon Instruments)
|
| Description |
Biological replicate: NoDox
|
| Data processing |
fluorescent ratios were log2 transformed, normalized and filtered in analogy to previous reports for mRNA expression microarrays (Bullinger et al. N Engl J Med 2004)
|
| |
|
| Submission date |
Aug 08, 2008 |
| Last update date |
Aug 11, 2008 |
| Contact name |
Lars Bullinger |
| E-mail(s) |
lars.bullinger@charite.de
|
| Phone |
+49-30-450-553111
|
| Organization name |
Charité
|
| Department |
Hematology, Oncology and Tumorimmunology
|
| Street address |
Augustenburger Platz 1
|
| City |
Berlin |
| ZIP/Postal code |
13353 |
| Country |
Germany |
| |
|
| Platform ID |
GPL7152 |
| Series (2) |
| GSE12394 |
murine MYC-dependent lymphoma cells: Dox vs. NoDox treatment |
| GSE12400 |
Analysis of MYC in murine lymphoma cell lines |
|
