Introduction to Recombinant Genetics- Biology 350

Mutagenesis

Mutagenesis Protocols

Radiation mutagenesis

Question:
If you mutate a plasmid, using chemical or radiation mutagenesis, how can create a situation where you limit the mutations to one segment of the output plasmid?

Nested Deletions

Linker scanning mutagenesis

Site-directed Mutagenesis

This method was first developed by Michael Smith, who was awarded a Nobel Prize in 1993 for this contribution. (URL)

Question:

After site-directed mutagenesis produces a mutant and wild type plasmid in the same bacteria, how could you select a purified mutant clone?

PCR Mutagenesis

Protocol

Tn5 Mutagenesis

Question:
Following transposon mutagenesis, how would you locate a transposon insertion into the genome?