Polymerase chain reaction-based site-directed mutagenesis using magnetic beads.
Thyroglobulin double mutants with various substitutions were obtained with a new polymerase chain reaction-based mutagenesis technique. Maximum length of megaprimer and efficiency of mutagenesis were improved by purification of single-stranded DNA, using the avidin-biotin interaction. This method might allow the construction of large libraries of DNA, mutated at different sites.
Fieschi
J
J
Laboratoire d'Ingénierie et Dynamique des Systèmes Membranaires, Institut de Biologie Structurale et de Microbiologie, CNRS, Marseille, France.
Niccoli
P
P
Camilla
C
C
Chames
P
P
Chartier
M
M
Baty
D
D
D001360:Avidin; D001483:Base Sequence; D017931:DNA Primers; D015723:Gene Library; D008280:Magnetics; D008863:Microspheres; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D013954:Thyroglobulin / Q000235:genetics*
D017931:DNA Primers; D001360:Avidin; D013954:Thyroglobulin