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The binary solvent system included mobile phase A (ACN/H2O, 60:40, v/v) and mobile phase B (IPA/ACN, 90:10, v/v), both containing 10 mM ammonium formate and 0.1% formic acid

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Extraction of replicated plasmid from bacterial cells Plasmid purification was performed using the Favorgen Endo-free Plasmid DNA Extraction kit (Favorgen, Taiwan), following the manufacturers protocol, with the quality of the extracted DNA assessed by electrophoresis on a 1% agarose gel before storage at -20 C