Ligation-independent cloning (LIC) is a form of molecular cloning that is able to be performed without the use of restriction endonucleases or DNA ligase. This allows genes that have restriction sites to be cloned without worry of chopping up the insert.
Protocol
- Design PCR Primers with LIC extension
- Perform PCR to amplify gene
- Purify PCR product
- Create 5' overhangs
- Incubate vector and PCR product to anneal
- Transform
References
Ligation-independent cloning Wikipedia(Text) CC BY-SA