PCR of large fragments

John Alsobrook ALSOBROOK at biomed.med.yale.edu
Fri Jul 17 15:25:50 EST 1992


In <1992Jul17.112214.202027 at uctvax.uct.ac.za> brknig01 at uctvax.uct.ac.za writes:

> 
> Hi netters.
> 
> If this keyboard hangs on long enough, I hope I can provide some info on PCR of
> long bits of DNA. 
> 
> A paper appeared in an early 1992 Nuc. Acids Res. about this very issue.
> In it, a compound called THESIT was prescribed to enhace the PCR reaction.

NO NO NO NO NO NO NO!!!!!!!!!!!!!!!!!

This is how methodological myths get started; the paper was NAR 20(3):623,
Ponce and Micol, "PCR amplification of long DNA fragments". If you actually
**READ** the paper, you'll see that they use "Thesit" in ALL of their buffer
conditions, *including* the standard Perkin-Elmer Cetus buffer. The *REAL*
difference in their conditions involves ELIMINATING KCl; they also tried
using Tricine rather than Tris. Of course, as I said in a previous posting,
they used a plasmid construct for their template. OK? OK!

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