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. The technique was developed in the early 1990s as an alternative to restriction enzyme/ligase cloning.[1] This allows genes that have restriction sites to be cloned without worry of chopping up the inserted gene of interest.[2][3][4][clarification needed]


  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


  1. ^ "Seamless Cloning | NEB". Retrieved 2020-01-23.
  2. ^ "Ligation Independent Cloning (LIC)". New England BioLabs (NEB). Retrieved 15 January 2016. CS1 maint: discouraged parameter (link)
  3. ^ "Get Your Clone 90% Of The Time with Ligation Independent Cloning". Bitesize Bio. Retrieved 2012-05-09. CS1 maint: discouraged parameter (link)
  4. ^ 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 readingEdit