Mutagenesis
From Wikipedia, the free encyclopedia
A process by which the genetic information of an organism is changed in a stable manner, either in nature or experimentally by the use of chemicals or radiation.
[edit] Types Of Mutation
The following are the types of Mutagenesis:
- PCR Mutagenesis
- Transposon Mutagenesis
- Site-directed mutagenesis
- Directed mutagenesis
- Insertional mutagenesis
- Targeted mutagenesis
[edit] PCR Mutagenesis
PCR mutagenesis is simple method for generating Site-directed mutagenesis. This method can generate mutations (base substitutions, insertions, and deletions) from double-stranded plasmid without the need for subcloning into M13-based bacteriophage vectors and for ssDNA rescue. The procedure involves a PCR reaction using a supercoiled plasmid vector as the template and two synthetic oligonucleotide primers containing the desired mutation with each complementary to the opposite strands of the vector. After PCR, the template (wild type) plasmid which is dam methylated in almost all E. coli is removed by digestion with Dpn I which is specific for methylated DNA. The mutated plasmid remains intact in the reaction.
