Ligation-independent cloning

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.[1][2][3]

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

Notes

  1. ↑
  2. ↑ "Get Your Clone 90% Of The Time with Ligation Independent Cloning". Bitesize Bio. Retrieved 2012-05-09.
  3. ↑ Haun, RS; Serventi, IM; Moss, J (1992). "Rapid, reliable ligation-independent cloning of PCR products using modified plasmid vectors". BioTechniques 13 (4): 515–8. PMID 1362067.

Further reading