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Which vector for cloning big PCR fragments? - (Jan/28/2005 )

blink.gif do you know a vector plasmid for cloning big pcr fragments? 15-20kb!


how did you get a PCR pdt that huge?? Is this a fusion of some sort?

You may have to use BACs for that size insert, and you have to cut it with a restriction enzyme and ligate it into the MCS of the BAC.


¡¡yes,BAC or cosmid can be considered.the same curiosity:how can you amplify such big DNA fragment by PCR? the tip will be quite helpful for us!


i used LATaq polymerase TAKARA inc.
they say you can amplify 35 kb. i arrived to 19kb. part of a natural plasmid in V. cholerae!thanks a lot