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For each mtDNA damage qPCR reaction, we used 15 ng DNA template, 0.4 M each of forward (5-TCT AAG CCT CCT TAT TCG AGC CGA-3) and reverse (5-TTT CAT CAT GCG GAG ATG TTG GAT GG-3) primers, nuclease-free water, and LongAmp Hot Start Taq 2 Master Mix (New England Biolabs), as described [108]

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When estrogen levels decline, gut integrity and microbiome balance often decline alongside it