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The cell pellet was resuspended in ice-cold 100 mM Tris, pH 7.6, centrifuged again, and finally resuspended in 1 mL of the same buffer

Once antibiotics are discontinued, TMAO levels often rise again

Opiates have long been used for a variety of medical conditions, with evidence of opiate trade and use for pain relief as early as the eighth century AD

7 Pick a verified colony and prepare competent cells as in steps 13

Following incubation, membranes were washed three times with TBST (5 min each) and incubated for 2 h at room temperature with alkaline phosphatase (AP)-conjugated secondary antibodies: goat anti-mouse (ab97020, Abcam 1:5000 dil.) or goat anti-rabbit (#ab6722, Abcam 1:3000 dil.), depending on the host species of the primary antibodies