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Fri 23 Jun, 2006 07:51 pm
Hi I am doing a Site Directed Mutagenesis and am wondernig why DNA LIGASE is not necsry after I do a PCR to amplify the dsDNA strands. Are the mutated prmiers not nicked one PCR is done, dont they need to be sealed to work--
in general does DNA not have to be ligated to "work".
Secondly: is DNA linear once it comes out of the PCR machine?
Little unclear on the question -- all PCR does is amplify short segments of DNA. You'd only need DNA ligase if you then want to try and integrate them back into a larger piece of DNA (be it a circular or a linear chromosome) -- and presumably the host cell carries its own ligase.
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