Use of In Vivo Assembly for High-efficiency Plasmid Construction

My doctoral research is focused on studying proteins that are secreted by bacteria. I specifically want to understand if some of these secreted proteins are important for causing human infections. A major challenge in working with recently isolated clinical bacteria is their antibiotic resistance, which hinders molecular genetic techniques that rely on antibiotic selection, such as gene deletions and genetic complementation.

This cloning protocol is faster and more cost effective since IVA eliminates the need for specialized enzymes and sequential enzymatic reactions for plasmid assembly before E.coli transformation. To begin, launch a specialized DNA analysis software on a computer system. Artificially assemble the desired plasmid.