title: Human kappa chain expression in a lambda 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 kappa 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.
authors.affiliation: Audie L. Murphy Memorial Veterans Hospital, San Antonio, TX.
authors.forename: K
authors.initials: K
authors.lastname: Lazaridis
authors.forename: H
authors.initials: H
authors.lastname: Dang
authors.forename: N
authors.initials: N
authors.lastname: Talal
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
chemical_list: D007145:Immunoglobulin kappa-Chains; D004247:DNA