LA-PCR ends?

rhubner at molbiol.ox.ac.uk rhubner at molbiol.ox.ac.uk
Fri Nov 25 17:20:38 EST 1994


hi you all,
 re:LA-PCR, is it known whether the products will be blunt (exo+ activity) or
should/can be used for TA-cloning approaches (terminal transferase activity)...
 A guy in our lab had problems with inverse PCR of a plasmid and I tried for the
first time the LA-PCR approach ("only" 4.4kb) and this worked extremely well...
 He seems to have had a frameshift with DynaZyme polymerase alone and attributes
this to terminal transferase activity. Hmmm, I really don't know HOW this could
happen on a _circular_ plasmid... Any explanations at hand? Trying several exo+
polymerases alone resulted in smears (yeah, it seems we join the club!)... we
sincerely hope that the joke has disappeared now!
 Thanks for any reply... it's GREAT to be with you all year 'round, like Xmas
EVERYday...
Have fun,
 Roland

ooops, I can not add this disclaimer bit from file... I'll try so from memory:
xxxnoxxx affiliation with xxxanyxxxx company, no xxxxx prejudice xxxxxx 
whatsoever, xxxxx no product xxxxxx...  



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