Seamless Cloning->LIC (ligation independent cloning)

brett brett at BORCIM.WUSTL.EDU
Fri Aug 16 16:05:40 EST 1996


>Dear Jeanne,
>
>
>You might also want to consider LIC (ligation-independent-cloning),which
>is described in the references below, as allowing directional cloning
>of PCR products without using ligations, restriction enzymes or T/A
>vectors.
>
>Basically, one adds an extra 11-12 bases to the 5' end of your
>PCR primers. After PCR, the PCR product ends are digested in the 
>presence of T4DNApol with dTTP or dATP which exposes the 11-12
>LIC site at the ends of your PCR product. This is *annealed*
>with the LIC vector at RT for about 30min and then transformed
>into e coli (NO ligations, NO restriction digests).

Maybe you could comment on the cost effectiveness of this method. As you
describe it, you'd be adding an aditional $20 in nucleotides per PCR just to
clone the thing. I have never had too much difficulty in standard RE cloning of
PCRs, so I'm not convinced of the utility of LIC.



Brett Lindenbach
    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu                             

"I own my own pet virus. I get to pet and name her." - Cobain




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