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Following DNA extraction, the mtDNA copy number in each sample was quantified by real-time polymerase chain reaction (PCR).
Protein in each sample was quantified using the BCA method.
Total DNA in each sample was quantified by OD260.
The absolute level of mRNA for the clock/functional genes in each sample was quantified.
Total protein in each sample was quantified using the BCA protein assay (Pierce, Rockford, IL).
The viral DNA in each sample was quantified by comparing the fluorescence profiles with a set of DNA standards.
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A specific standard curve was performed in parallel for each gene, and each sample was quantified in duplicate.
Then, the concentration of the protein in the extracted tissue of each sample was quantified using the Bio-Rad Protein Assay kit (BioRad Laboratories), with bovine seroalbumin (BSA) as the standard protein.
Each of the immunoprecipitations was repeated independently once, and each sample was quantified in triplicate.
For every promoter fragment analyzed, each sample was quantified in triplicate from 2 to 5 independent immunoprecipitations. On Q-PCR from animals are represented the mean of five independent experiments performed on triplicate, on Q-PCR from cells is shown one representative experiment done on triplicate.
Each sample was quantified in duplicate.
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