A rapid PCR method of screening for small mutations.
We report a modified method of screening for point mutations using a PCR approach based upon the sensitivity of PCR to the 3' terminus of the primer. This method provides a sensitive screen when using either plasmid DNA or bacterial cell lysates. We have optimized the technique for general use to allow rapid screening of mutants with good discrimination. Unlike previous similar methods, this technique has no inherent limitation in primer design on the 3'-terminal base chosen.
Major
J G
JG
Hybritech, Inc., San Diego, CA 92086.
D001483:Base Sequence; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic; D001618:beta-Lactamases / Q000235:genetics
D004269:DNA, Bacterial; D001618:beta-Lactamases