|
| Status |
Public on Oct 01, 2015 |
| Title |
MDAMB231-negative_control-rep3 |
| Sample type |
RNA |
| |
|
| Source name |
MDA-MB-231 cells, mir-184 mimic negative control, 48 hours
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MDA-MB-231
gender: female
tissue: breast
transfection: mir-184 mimic negative control, 48 hours
|
| Treatment protocol |
MDA-MB-231 cells were seeded in 6 well plates at a cell density of 1.8 x 105 cells/ well. The next day, miRIDIAN miRNA mimics (Dharmacon) were transfected into cells using Lipofectamine 2000 (Invitrogen) according to the manufacturers protocol for transfecting siRNA. Mimics (Final concentration of 50nm) was mixed with 1% lipofectamine 2000 (v/v) diluted in Opti-MEM transfection medium (Invitrogen) and incubated for 20 minutes. The mimics were added dropwise onto cells in growth medium at a final concentration of 50nm. Fresh medium was replaced 24hours post transfection.
|
| Growth protocol |
MDA-MB-231 cells were cultured in RPMI 1640 media supplemented with 10% FCS at 37°C, 5% CO2. Cells were subcultured every 3 to 4 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using Trizol reagent (Invitrogen) using the manufacturers protocol and further ethanol precipitated to remove salt contaminants. The RNA was quantified using the Nanodrop 1000 spectrophotometre and the integrity of the RNA was determined using the Agilent 2100 bioanalyser. (Agilent Technologies, Santa Clara, CA)
|
| Label |
Biotin
|
| Label protocol |
RNA samples were reverse transcribed into biotinylated cRNA at the Ramaciotti Centre for Gene Function Analysis at the University of New South Wales (UNSW, Sydney, NSW, Australia), following manufacturers instructions.
|
| |
|
| Hybridization protocol |
Labeled, fragmented cDNA hybridized to Affymetrix Human Gene 1.0 ST Gene Arrays at the Ramaciotti Centre for Gene Function Analysis at the University of New South Wales (UNSW, Sydney, NSW, Australia), following manufacturers instructions.
|
| Scan protocol |
Arrays were scanned at the Ramaciotti Centre for Gene Function Analysis at the University of New South Wales (UNSW, Sydney, NSW, Australia), following manufacturers instructions.
|
| Description |
MDA-MB-231 cells, mir-184 mimic negative control, 48 hours
|
| Data processing |
Data was RMA normalised using NormalizeAffymetrixST GenePattern module from the Peter Wills Bioinformatics Centre's GenePattern Server: http://pwbc.garvan.unsw.edu.au/gp
|
| |
|
| Submission date |
Oct 10, 2014 |
| Last update date |
Oct 01, 2015 |
| Contact name |
Daniel Roden |
| E-mail(s) |
[email protected]
|
| Organization name |
Garvan Institute
|
| Street address |
384 Victoria Street
|
| City |
Sydney |
| State/province |
NSW |
| ZIP/Postal code |
2010 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6244 |
| Series (2) |
| GSE62259 |
MicroRNA profiling of the developing mammary gland identifies miR-184 as a candidate breast tumour suppressor gene (Affymetrix) |
| GSE62261 |
MicroRNA profiling of the developing mammary gland identifies miR-184 as a candidate breast tumour suppressor gene |
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