QuickChange Site-Directed Mutagenesis
Site-directed mutagenesis is a powerful technique that permits the introduction or deletion of any number of basepairs in a plasmid. Agilent sells a very expensive but useful kit for site-directed mutagenesis:
- QuickChange Lightning Kit – for introducing mutations at a single site.
- QuikChange Lightning Multi Site-Directed Mutagenesis Kit – for introducing multiple mutations simultaneously. Make sure to get academic prices!
Contents
Section titled “Contents”1. Notes
Section titled “1. Notes”- Inverse PCR Mutagenesis is a very easy technique for introducing mutations in your plasmid and does not require any kits. Large deletions are much easier to perform. Also, there’s no magic buffer mumbo-jumbo. I strongly recommend you use this method for routine mutagenesis
- Enzymes from this kit are stored in -20C
- Cells are stored in -80C, although using standard lab DH5a cells works just as well as the fancy Agilent cells.
2. Introducing a Single Mutation
Section titled “2. Introducing a Single Mutation”Please read the manual: quickchange_lightning_mutagenesis_kit.pdf 
3. Introducing Multiple Mutations
Section titled “3. Introducing Multiple Mutations”Please read the manual: agilent_multisite_kit.pdf 