Article
Simple biological assay for the error rate upon cloning of synthetic oligodeoxyribonucleotides.
BioTechniques - 1 Jul 1990
Mandecki W, Shallcross M A, Kavanaugh T J
Abstract excerpt
A method was developed for determination of the rate of undesired point mutations upon cloning of synthetic DNA. The method relies on cloning of an oligonucleotide(s) into the E. coli alkaline phosphatase gene inactivated due to a small deletion within the active site. The oligonucleotide adds back the deleted sequence, but simultaneously introduces a missense mutation at a critical position. The activity of the...
Topics
- Alkaline Phosphatase
- Amino Acid Sequence
- Bacterial Proteins
- Base Sequence
- Biological Assay
- Cloning, Molecular
- Genes, Synthetic
- Genetic Vectors
- Molecular Sequence Data
- Mutation
- Oligodeoxyribonucleotides