"RediLoad" PCR loading buffer recipe

John Ladasky jladasky at pmgm.Stanford.EDU
Fri Feb 13 09:26:02 EST 1998


In article <dneece-1102981047110001 at klbg433c.tamu.edu>,
David Neece <dneece at startel.net> wrote:
>I'm looking for the recipe that is the equivalent of Research Genetics'
>"RediLoad" PCR loading buffer.  This buffer/dye is added to the reaction
>before amplification and serves as a loading buffer/dye when running the
>gel.  I've been told that there was a publication out a few years back
>that compared several such buffers, and that the RediLoad recipe is
>contained in this reference.  I'd greatly appreciate it if anyone could
>supply me with the recipe or reference.

	I don't know anything about Research Genetics or Rediload buffer.
However, I find that I can add a solution of sucrose, cresol red, and 
tartrazine yellow directly to my PCR.  This does not seem to affect the 
reaction conditions much, and it allows me to load the PCR product direct-
ly onto my agarose gel for analysis without mixing it with a separate 
loading buffer.  

	The recipe for 20X PCR compatible loading buffer is as follows:

	- 30% (w/v) sucrose
	- 0.2% (w/v) cresol red
	- 0.3% (w/v) tartrazine yellow
	- sterilize by autoclaving. I have also tried UV irradiation to
	  inactivate any contaminating DNA with no apparent side-effects

	I got this idea from a recent Biotechniques article.


-- 
Rainforest laid low.
"Wake up and smell the ozone,"
Says man with chainsaw.					- John Ladasky



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