Ligation Independent Cloning literally says no need Ligation, but it still needs ligase to anneal the fragment to vector. And the Protocol used T4 DNA Ligase for anneal process. What does this mean "Ligation Independent"? SLIC Protocol

  • $\begingroup$ Please provide a proper scientific reference for the method — your link is not to such, and in any case is dead. You write that ligase is needed in the annealing step, but this is not true. What makes you think that? $\endgroup$
    – David
    Apr 21 at 9:02
  • $\begingroup$ @David Ligase works for annealing the Cohesion end. $\endgroup$ Apr 21 at 17:34
  • $\begingroup$ Annealing means hybridization by complementary base-pairing. It involves non-covalent hydrogen-bonding. Ligation is a separate process in which the hybridized species might be covalently joined. In LIC the long overhangs keep the recombined together without ligation being needed in vitro. This happens in and by the transformed bacteria. $\endgroup$
    – David
    Apr 21 at 22:12


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