Overlapping extensions generate site-specific mutagenesis experiments
Four primers are required for site-specific mutagenesis by the overlapping extension method (see Fig. 13-4) (Higuchi et al. 1988, Hetal. 1989). This experiment
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Efficient and rapid sentinel mutagenesis in the same tube using large-primer PCR
The large primer method was originally established by KAMMANN et al. (1989) and the current method has been modified by many researchers including Sarkat and So
Screening experiments of small quantities of bacterial clones by hybridization method
Bacterial clones on the main agar plate and on a nitrocellulose filter membrane or nylon membrane covering the surface of another agar plate can be localized, a
In vitro mutagenesis using double-stranded DNA as a template: mutant selection experiment with DpnⅠ
This scheme and Scheme 4 use two oligonucleotides and high-fidelity polymerase to guide DNA synthesis using denatured plasmid DNA as a template. In this scheme,
Selection of plasmid vectors to construct expression libraries
There are three methods for detecting bound antibodies: radiochemical screening, colorimetric screening, and chemiluminescent screening, the advantages and disa
Exonuclease III digestion produces multiple sets of nested deletion mutants
The nested deletion mutagenesis method, in which multiple oligonucleotides are progressively deleted from one or the other end of the target DNA, has been used
Cross-reactive antibody removal assay in antisera
This protocol describes a method for removing antibodies reactive with bacterial-encoded proteins from polyclonal antisera by co-warming the antiserum with bact
Affinity chromatography assay for removal of cross-reactive antibodies
This section describes a method for removing components of crude immunoglobulin that react with bacterial-encoded proteins using a matrix that binds bacterial p
Antibody assay for removal of cross-reactivity in antisera
The methods described in this section are only for the preparation of antisera containing relatively low-titer anti-E. coli antibodies. If the treated antiserum