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Quantitative real-time PCR was performed with either 2.5 L diluted cDNA (12.5 ng RNA), a 10 L Kapa SYBR FAST qPCR Master Mix Universal (KapaBiosystems, Wilmington, MA, US), or 9 L diluted cDNA (25 ng RNA) and 1 L predeveloped TaqMan Gene Expression Assays were mixed in 10 L TaqMan Gene Expression Master Mix (Thermo Fisher Scientific, Waltham, MA, US), on a Bio-Rad CFX96 Touch Real-Time PCR Detection System (Bio-Rad Laboratories, Hercules, CA, US)

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For circadian and melatonin research, timing of administration relative to light-dark cycles is a methodologically important variable, given Epithalon's interaction with pineal transcriptional machinery that is itself circadian-regulated

N., & Adams, G