Ligation-independent cloning


Ligation-independent cloning is a form of molecular cloning that is able to be performed without the use of restriction endonucleases or DNA ligase. The technique was developed in the early 1990s as an alternative to restriction enzyme/ligase cloning. This allows genes that have restriction sites to be cloned without worry of chopping up the inserted gene of interest.

Protocol

  1. Design PCR Primers with LIC extension
  2. Perform PCR to amplify gene
  3. Purify PCR product
  4. Create 5' overhangs
  5. Incubate vector and PCR product to anneal
  6. Transform