|
| Status |
Public on Dec 31, 2010 |
| Title |
MB231 transfected with Dmimic, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
MDA-MB-231 cells grown in 6 well dishes 48 hrs post-transfection
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MDA-MB-231
|
| Treatment protocol |
MDA-MB-231 cells (600,000 cells/well) were reverse transfected in 6 well dishes with either AllStar negative control, Qmimic, or Dmimic at 10 nM final concentration using Lipofectamine 2000 (10.5 ul/well),
|
| Growth protocol |
MDA-MB-231 cells were grown in T75 flasks containing RPMI 1640 basal medium + 10% Fetal Bovine Serum at 37C with 5% CO2. Cells at 80-90% confluency were used in transfections.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
At 48 hrs post-transfection, total RNA was extracted from each well using Qiagen's miRNeasy extraction kit according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
RNA quality ws assessed on an Agilent Bioanalyzer. Total RNA (1 ug) was reverse transcribed with T7-oligo(dT) primer and labeled with biotin using Affymetrix One Cycle Target Labeling kit following the manufacturer's protocol.
|
| |
|
| Hybridization protocol |
cRNA was hybridized for 16 hrs at 45C (60 rpm). GeneChips were washed and stained in the Affymetrix FS450 fluidics station.
|
| Scan protocol |
GeneChips were scanned using a Affymetrix GeneChip 3000 Scanner. Data was collected using Affymetrix GCOS software.
|
| Description |
Gene expression data of total RNA extracted from MB231 cells 48hrs post-transfection
|
| Data processing |
Expression values were normalized using Robust Multichip Averaging (RMA). Analyses were performed with Bioconductor packages AFFYGUI and LIMMA on a R environment.
|
| |
|
| Submission date |
May 14, 2010 |
| Last update date |
Dec 31, 2010 |
| Contact name |
Mark Mackiewicz |
| E-mail(s) |
mackiewiczm@mail.nih.gov
|
| Phone |
301-594-6044
|
| Fax |
301-480-8878
|
| Organization name |
National Institutes of Health
|
| Department |
National Cancer Institute
|
| Lab |
Genetics Branch
|
| Street address |
37 Convent Drive
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (2) |
| GSE21832 |
Identification of the receptor tyrosine kinase AXL in triple negative breast cancer as a novel target for the human miR-34a microRNA (gene expression) |
| GSE21834 |
Identification of the receptor tyrosine kinase AXL in triple negative breast cancer as a novel target for the human miR-34a microRNA |
|
