"SMEAR" BUT NO BANDS WHEN DOING "LONG" PCR USING KLENTAQ1 WITH PFU

David Sang-shin Lee davidlee at umich.edu
Mon May 16 18:28:11 EST 1994


Christian Beltinger (betinger at pobox.upenn.edu) wrote:

> RE:  "SMEAR" BUT NO BANDS WHEN DOING "LONG" PCR USING KLENTAQ1 WITH PFU	 

> 		Consistently, I get these surprising results when running out 25-50 ul on
> an 0.8% agarose gel:  Instead of discrete bands there is an intense
> homogenous smear from the well down to the dye front , even in samples
> without DNA template!

> 	Does anyone have an explanation for this phenomenon and a suggestion how
> to avoid it?

I use Taq/Vent, and run my products on an acrylamide gel, stained with 
ethidium bromide afterwards.  I get a smear every time, even when I have 
a discrete product.  Perhaps this is due to the BSA in the buffer?

-------------------------------------------------------------------
David S. Lee  (davidlee at umich.edu)                   All opinions
HR6028, Kresge Hearing Research Institute             expressed
University of Michigan                                 here are
Ann Arbor MI  48109-0506                                mine!



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