PCR Subcloning with restriction sites

John Thompson jrt at home.com
Sat Apr 1 21:44:29 EST 2000


Look at the cut, ligate and recut specs in the catalogs (at least NEB
quotes these).  They vary widely for different enzymes.  Don't have a
catalog handy but you'll see something like: 10X overdigestion can be
90% ligated and recut vs. 100X overdigestion can be 95% ligated and
recut.  

John Thompson
Merck Research Labs


Tom Knight <tk at pasteur.ai.mit.edu> wrote:

>There seems to be a lot of "lore" about which enzymes are good and bad
>for subcloning.  Would someone care to enlighten us on which are the
>"good" and which are the "bad" enzymes?  The catalogs seem to only
>sell the good ones.... ;-)





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