Molecular Evolution: Random Mutagenesis
- Three ready-to-go kits for random mutagenesis of a gene of interest
- Accompanied by detailed manuals and background information
Within three billion years of evolution, nature has produced a plethora of proteins simply by repeated cycles of random mutagenesis followed by in vivo
selection. This example of natural evolution has guided researchers to develop strategies for in vitro
permutation of genes and proteins. Among a variety of strategies, three powerful techniques have emerged:
Mutagenesis by dNTP Analogs
The method achieves mutagenicity of up to 20% and is based on incorporation of mutagenic dNTP analogs into an amplified DNA fragment. The analogs are eliminated by a second PCR in the presence of the four natural dNTPs, leaving highly mutated DNA ready for further investigation.
Mutagenesis by Error-Prone PCR
Mutagenesis is performed by a PCR reaction under modified conditions that induce an increased error-rate of the DNA-polymerase. Simply run the PCR protocol provided in the manual and achieve mutagenicity in the range of 0.6-2.0% in a single PCR reaction!
Mutagenesis by DNA Shuffling
DNA shuffling generates libraries by random fragmentation of one gene or a pool of related genes, followed by reassembly of the fragments in a self-priming PCR reaction. The mutagenicity is similar to error-prone PCR but DNA shuffling allows recombination of sequences from different, related genes.