|
| Status |
Public on Aug 08, 2012 |
| Title |
NIH-3T3_Risc-transfected_48h_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
NIH-3T3_Risc-transfected_48h
|
| Organism |
Mus musculus |
| Characteristics |
cell line: NIH-3T3
cell type: murine fibroblast cells
transfectant: RISC-free siRNA (control)
|
| Treatment protocol |
Cells were transfected with 25nM RNA complexed to DharmaFECT 1 and analysed after 48 hours.
|
| Growth protocol |
NIH-3T3 fibroblasts were grown in RPMI supplemented with 10% FCS, 2mM L-glutamine, 0.25U/ml penicillin and 100U/ml streptomycin at 37C 5%CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the Qiagen miRNeasy kit as described by the manufacturer.
|
| Label |
biotin
|
| Label protocol |
Total RNA was converted to cDNA using the GeneChip WT cDNA Synthesis and Amplification Kit (Affymetrix) and biotinylated using the GeneChip® WT Terminal Labeling Kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 5.5 ug of sscDNA were hybridized for 16 hr at 45C on Affymetrix GeneChip ® Mouse Gene 1.0 ST Array.
|
| Scan protocol |
GeneChips were scanned using the Agilent Technologies Micro Array Scanner #G2505B.
|
| Description |
Risc_3
gene expression data from cells transfected with control RNA.
|
| Data processing |
The data were analyzed with Partek Genomics Suite version 6.5 using Affymetrix default analysis settings and RMA as normalization method. Based on the log 2 value of negative control probes of each chip, the background fluorescence was calculated in StatPlus and used to eliminate the microarray's background noise.
|
| |
|
| Submission date |
Jan 05, 2012 |
| Last update date |
Aug 09, 2012 |
| Contact name |
Dominik Andreas Rückerl |
| E-mail(s) |
dominik.ruckerl@manchester.ac.uk
|
| Organization name |
The University of Manchester
|
| Department |
Faculty of Biology, Medicine and Health
|
| Street address |
Oxford Road
|
| City |
Manchester |
| ZIP/Postal code |
M13 9PT |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE34873 |
Experimental identification of miR-378-3p targets by overexpression and silencing in murine NIH-3T3 fibroblasts |
|
