Male and female B6C3F1/N mice were exposed to 0, 200, 600, and 2,000 mg/kg GBE by corn-oil gavage, 5 days a week for 2 years.
Extracted molecule
total RNA
Extraction protocol
miRNA extraction was performed using MirVANA miRNA Isolation Kit (Life technologies, Carlsbad, CA) and RNA integrity was measured with Bioanalyzer (Agilent Technologies, Santa Clara, CA).
Label
biotin
Label protocol
miRNA expression analysis was conducted using Affymetrix GeneChip® miRNA 3.0 Array (Affymetrix, Santa Clara, CA) following the Affymetrix hybridization protocols. According to the manufacturer’s protocol, the Affymetrix FlashTagTM Biotin HSR labeling (Affymetrix, Santa Clara, CA) was used to label 200 ng of total RNA.
Hybridization protocol
Each labeled sample was hybridized for 16 hours at 48°C in a rotating hybridization oven using the Affymetrix Eukaryotic Target Hybridization Controls (Affymetrix, Santa Clara, CA) and protocol. Array slides were stained with streptavidin/phycoerythrin utilizing a double-antibody staining procedure and then washed for antibody amplification according to the GeneChip Hybridization, Wash and Stain Kit user manual.
Scan protocol
Arrays were scanned in an Affymetrix Scanner 3000 and data was obtained using the GeneChip® Command Console Software (AGCC; Version 3.2) and Expression Console (Version 1.2).
Description
miRNA Expression Profile
Data processing
The raw miRNA data were normalized using the Robust Multichip Analysis (RMA) approach
Unique microRNA Alterations in Hepatocellular Carcinomas Arising Either Spontaneously or due to Chronic Exposure to Ginkgo biloba Extract (GBE) in B6C3F1/N Mice
