Surf Wiki
Save to docs
general/cloning

From Surf Wiki (app.surf) — the open knowledge base

Ligation-independent cloning

Ligation-independent cloning (LIC) is a form of molecular cloning that can 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 to be cloned without the requirement of a restriction site for cloning that is absent from the gene insert. LIC uses long complementary overhangs on the vector and the DNA insert to create a stable association between them.

Steps in Procedure

  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

References

References

  1. "Seamless Cloning {{!}} NEB".
  2. "Ligation Independent Cloning (LIC)". New England BioLabs (NEB).
  3. "Get Your Clone 90% Of The Time with Ligation Independent Cloning". Bitesize Bio.
  4. (1992). "Rapid, reliable ligation-independent cloning of PCR products using modified plasmid vectors". BioTechniques.
  5. (2020). "Bacterial genetics and genomics". CRC Press.
Info: Wikipedia Source

This article was imported from Wikipedia and is available under the Creative Commons Attribution-ShareAlike 4.0 License. Content has been adapted to SurfDoc format. Original contributors can be found on the article history page.

cloning
Want to explore this topic further?

Ask Mako anything about Ligation-independent cloning — get instant answers, deeper analysis, and related topics.

Research with Mako

Free with your Surf account

Content sourced from Wikipedia, available under CC BY-SA 4.0.

This content may have been generated or modified by AI. CloudSurf Software LLC is not responsible for the accuracy, completeness, or reliability of AI-generated content. Always verify important information from primary sources.

Report