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The cDNA was then subjected to quantitative real-time PCR (10 ng per reaction) carried out with TaqMan Universal PCR Master Mix (Life Technologies, Carlsbad, CA, USA) using the 7300 Real-Time PCR System (Life Technologies) with pre-developed TaqMan probe/primer combinations for UCP-1 (Mm00494069_m1) and glyceraldehyde-3-phosphate dehydrogenase (Mm99999915_g1) optimized by the manufacturer
