Human κ chain expression in a λ phage vector: methods of isolating amplified cDNA affects cloning efficiency

Abstract

Three common methods of isolating amplified DNA were evaluated for their effects on cloning efficiency in a phage vector designed to express human κ chain. The “glass milk” technique gave higher cloning efficiency and protein expression than phenol-chloroform extraction or microfiltration. This shows that the quality of amplified cDNA should be considered when studying the human antibody repertoire using this vector system.