Plasmid DNA extraction was conducted using a previously reported method [20 (link)]. The PureLink HiPure Plasmid DNA Purification Kit (Invitrogen, Carlsbad, CA, USA) was employed, following the manufacturer’s instructions. The extracted plasmid DNA solution was quantified using a Qubit Flex Fluorometer (Thermo Fisher Scientific, Wilmington, DE, USA) based on fluorometry. To verify the integrity of the plasmid DNA, electrophoresis was performed in 2% agarose gel. Subsequently, the samples were stored at −80 C until further use.
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