Experiments on the preparation of denaturing polyacrylamide gels
In sequencing, 4 series of DNA fragments are analyzed by electrophoresis on a thin polyacrylamide gel under denaturing conditions. This protocol describes the p
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Chemical sequencing assay experiment
The importance of chemical sequencing has always been demonstrated by obtaining sequences of oligonucleotides, functional analysis of transcriptional regulatory
Experiments on the preparation of polyacrylamide gels containing formamide
A common problem in DNA sequencing is the compression of electrophoretic bands due to the presence of DNA dipolar structures. A simple and straightforward solut
Cycle sequencing: double deoxy sequencing experiments using PCR and primer end-labeling
The key to the success of this method is the purity of the template DNA. Contamination with agarose, salt, and protein can lead to premature termination or susp
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
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 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
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
Oligonucleotide-directed single-stranded DNA mutagenesis experiments
This paper describes a classic protocol consisting of the double primer technique of Zoller and Smith (1984,1987) combined with Kunkel's mutant yield enrichment