One-Step Bacterial Artificial Chromosome (BAC) Modification: Transformation of the BAC Host with the RecA Vector

Abstract

This protocol outlines the steps for introducing the RecA plasmid into bacterial artificial chromosome (BAC) host cells, and their preparation for subsequent transformation with the reporter plasmid for one-step BAC modification. BAC host cells are rendered chemically competent and transformed with the RecA plasmid, and transformants are selected for tetracycline resistance to ensure the presence of the RecA marker.