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Both the Tet-inducible vector pRMC2 (Addgene, plasmid 68940) 70 and the mKate insert were digested with KpnI-HF and EcoRI-HF (New England Biolabs), gel purified using a QIAquick PCR purification kit (Qiagen), and ligated using T4 DNA ligase (New England Biolabs) at 16 °C for 16 h

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For researchers familiar with semaglutide appetite suppression timelines , the GLP-1 component of CagriSema follows the same general pattern