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
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Sampling and DNA sequencing
DNA sequences can be enzymatically or chemically degraded to produce a series of sets of DNA fragments that can be resolved by thin-layer denaturing polyacrylam
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
Chemical sequencing assay experiment
The importance of chemical sequencing has always been demonstrated by obtaining sequences of oligonucleotides, functional analysis of transcriptional regulatory
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
Electroshock transformation experiments in Escherichia coli
Electroshock transformation experiments in E. coli can be used for: easier and faster transformation.
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
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
USE mutation experiment
In this method, two oligonucleotide primers hybridize to the same strand of the DNA duplex of a denatured recombinant plasmid. One primer (mutagenic primer) car