Screening experiments of intermediate cell clones by hybridization method
This method is applicable to bacterial clones of any size, but small clones give the clearest results. Small clones produce sharp hybridization signals and are
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Uracil-containing single-stranded phage M13 DNA preparation experiments
The classical Kunfcel oligonucleotide-directed mutagenesis method utilizes the selective action of uracil DNA glycosylase in E. coli to specifically screen out
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
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,
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
Single-strand conformational polymorphism and heteroduplex analysis methods for mutation detection experiments
The purpose of sinister mutation testing is mainly for figuring out the molecular mechanisms of human hereditary diseases in order to effectively target these d
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
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
Screening of fixed-point mutagenesis recombinant cloning experiments by hybridization with radiolabeled oligonucleotides
This protocol mainly describes the screening of recombinant phage M13 clones, whereas one option is to screen bacterial clones containing phage particles. Final