The Licase™ cloning method results from the work of Dr. Philippe Valadon at Antibody Design Labs while exploring novel methods to accelerate the step of DNA ligation. The principle relies on a thermal switch that controls a 3′->5′ exonuclease activity to generate single-stranded DNA overlaps between DNA fragments. In the first phase, the exonuclease activity rapidly creates complementary overhangs on the DNA fragments. After switching the temperature to 75°C for a few seconds, the exonuclease activity is replaced by an annealing activity which results in the ultra-fast assembly of the DNA fragments through homologous recombination. The reaction mixture is immediately ready for bacterial transformation. Since the DNA linking event is based on sequence homology, the assembly reaction is highly specific, producing a very high cloning success rate, sometimes near 100%. The very short ligation time is changing markedly the flow through of molecular cloning at the bench.
Advantages of Licase™ Cloning
Super fast (5 s).
Very high success rate.
High-efficiency competent bacteria unneeded.
No positive control required.
Works for simple cloning, multi-fragment assembly, mutagenesis.
Faster and cheaper than competition.
PUB011 – Rev170415
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