7 protocols
AccessionType
image_acquisition
labeling
hybridization
bioassay_data_transformation
RMA normalisation, as in the Nimblegen user's guide.
bioassay_data_transformation
The normalized file contains gene expression values produced using the Robust Multichip Average (RMA) algorithm.
nucleic_acid_extraction
Total RNA was extracted from each well of transfected cells using Trizol reagent (Invitrogen). RNA was then pooled from 3 independent transfection replicates per condition in an equimolar manner. Nimblegen performed the double stranded cDNA synthesis and hybridization step.
grow
COV434 cells were grown in DMEM-F12 medium supplemented with 10%FBS and 1% Penicilin-Streptomycin, and plated at 40% confluence in 6-well plates 24h before transfection. Cells were transfected with the Lyovec (Invivogen) reagent using 25ug plasmid DNA per well.