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Does anyone have experience with LONG PCR?

Eric C. Anderson anderson at pharmdec.wustl.edu
Mon Sep 12 10:02:14 EST 1994


In article <kamin_johnson-0909941524330001 at bongo.cellbio.duke.edu>,
kamin_johnson at cellbio.duke.edu (kamin johnson) wrote:
> > We are attempting to amplify a 16 kb fragment and would like to hear
> > about other's experience with using technique, buffer recipes,
> > protocols, and kits tried.
> > Any or no success using this, we'd like to know.  

>    In a recent issue of TIBS (I think, one of the Trends... issues anyway)
> there was a discussion of the "long and accurate" PCR technique.  The
> point was that the technique seems to require one to follow the protocol
> of Barnes rigidly.  The protocol was given in the issue.

i too reccomend Wayne Barnes' protocol for Long distance PCR.  while the
enzyme he designed specifically for LD PCR is not commercially available,
Stratagene's Taq Extender PCR additive was developed in conjunction with
Wayne, and, while they didn't get 16kb products, they can consistently
amplify 10kb with very little atifactual products.

good luck,
eric

-- 
*************************************
*Of course it's not true...         *
*But let's make the bastard deny it!*
*                        LBJ (1948) *
*************************************
eric c. anderson                                
anderson at pharmdec.wustl.edu
dept. of molecular bio. and pharm.               (314)362-3963 (lab)
washington univ. school of medicine              (314)862-2435 (home)
660 s. euclid box 8103                           (314)362-7058 (FAX)
st. louis, mo 63110



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