dose: 0 mg/kg DB[a,h]A tissue: part of liver gender: male age: 10 weeks genotype/variation: MutaMouse (transgenic mouse strain 40.6) date of exposure: January 24- February 22 time post exposure: 3 days
Treatment protocol
10-week old male Muta™Mouse (i.e., transgenic mouse strain 40.6) were dosed daily via oral gavage for 28-days with DB[a,h]A dissolved in olive oil (6.25, 12.50 and 25 mg/kg body weight/day). Each dose group contained 5 animals; 5 animals were also dosed with a vehicle (olive oil) control. Following a 3 day expression period after the last treatment, mice were euthanized with isofluorane followed by cervical dislocation. Liver was then excised, flash-frozen in liquid nitrogen, and stored at -80ºC until use.
Extracted molecule
total RNA
Extraction protocol
Total RNA containing the small fraction was isolated from each sample using the miRVana miRNA Isolation Kit (Ambion), Streetsville, ON, Canada). Quality was confirmed using the Agilent 2100 Bioanalyzer and RNA Nano 6000 chips.
Label
Cyanine 3-pCp
Label protocol
Total RNA samples were labelled using the Agilent miRNA Complete Labelling and Hybridisation Kit (Agilent Tech, Mississauga, ON, Canada).
Hybridization protocol
Hybridisations were performed in Agilent SureHyb chambers for 20 hours at 55C and washed according to manufacturer's instructions.
Scan protocol
Arrays were scanned at a resolution of 5 mm using an Agilent G2505B scanner.
Data processing
The log2 of median signal intensites were cyclic lowess normalized and the technical replicates were averaged using the median.
