7 protocols
AccessionType
normalization data transformation protocol
RMA-EXON-CORE-DABG protocol HuEx-1_0-st-v2.r2.pgf HuEx-1_0-st-v2.r2.dt1.hg18.core.mps ID_REF = VALUE = RMA signal estimates
array scanning protocol
hybridized microarrays were scanned on GeneChip Scanner 3000 (Affymetrix, P7N 00-00212)
hybridization protocol
fragmented and labelled cDNA was hybridized to GeneChip® Human Exon 1.0 ST (Affymetrix,900694) at 45°C for 16 hours, washing and staining was done using GeneChipHybridization Wash and Stain kit (Affymetrix, 900720)
labelling protocol
sense strand cDNA was prepared with Ambion WT Expression Kit (Life Technologies, 4411973), fragmentation and biotin labelling was done with GeneChip WT terminal labelling kit (Affymetrix, 900670)
growth protocol
Flp-In T-REx 293 cells were stably transfected with a plasmid encoding AGO1 N-terminally tagged with ProteinA-TEV protease cleavage site-Hisx6. Expression of tagged AGO1 is tetracycline inducible. For the purpose of this microarray analysis AGO1 expression was not induced. Cells were grown in Dulbecco’s modified Eagle medium (DMEM) with high glucose, supplemented with 10% fetal bovine serum at 37°C under humidified air with 5% CO2
nucleic acid extraction protocol
total RNA was izolated 48 hours post transfecton with TRIZOL (Life Technologies) according to manufacturers instructions; sample integrity was confirmed on Bioanalyzer and samples were further processed by the Source BioScience UK Limited
sample treatment protocol
Flp-In T-REx 293-PTH-AGO1 cells were transfected with Lipofectamine 2000 (Life Technologies) and miR-92a (mA/ZEN/mCmAmGmGmCmCmGmGmGmAmCmAmAmGmUmGmCmA mAmU/ZEN/mA; IDT; 6.25nM) or control inhibitor (mC/ZEN/mGmCmGmAmCmUmAmUmAmCmGmCmGmCmAmAmUmAmUmGmG/ZEN/mU; IDT; 6.25nM)