Fidelity of LA-PCR

Brian Desany bd033054 at condor.mbcr.bcm.tmc.edu
Wed Jan 3 13:48:51 EST 1996


Does anybody know how accurate BMB's expand system is compared to
Barnes's Klentaq1/Pfu formulation?  I cloned a 2.7kb gene with
Expand and found a mistake (2 independent products had one base 
different from each other, so its not a polymorphism of the
published sequence) in the first 300bp I sequenced.  I hope this is
a statistical fluke! I was hoping to be able to use this technology to
clone ORFs with convenient flanking restriction sites built into the
primers, but if I have to sequence multiple long clones every time
it becomes more trouble than its worth.  Should I switch over to 
the klentaq/pfu mix (which I know is even more accurate than pfu
which is billed as being very accurate) or is this approach just
not feasible/too risky?  Perhaps I could get some tips on maximizing
fidelity? Thanks-
-Brian.





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