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However, a 2025 review noted only 3 RCTs, calling for larger trials (Nguyen et al., 2025)

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After the initial denaturation step (7 min, 95 C), the qRT-PCR program was assembled (7 min, 95 C) by 40 repetitive cycles containing a denaturation step for 15 s at 95 C followed by an annealing extension for 60 s at 60 C

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