PCR and loading solution

WSchick at aol.com WSchick at aol.com
Sun Oct 27 13:41:39 EST 1996


In a message dated 96-10-20 14:48:35 EDT, melansond at am.abru.cg.com
(MELANSOND) writes:

<< 
 i find it hard to believe that one can add the dye and sucrose to the PCR
 reaction mixture without severely inhibiting the reaction, however, there
are
 many things I don't know. So if this truly is possible I would like to hear
 about it as well.  In the meantime...  I would suspect that one should add
the
 loading dye to the PCR reaction tube AFTER the reaction has taken place,
 immediately before loading the gel.  The concentrations depend on the
 concentration of the stock loading dye (10 X, 6X etc), but one would ideally
 like to add 1unit of dye / 4 units of reaction volume, to achieve a final
 concentration ratio of 1:5. >>


The "rapid cyclist" web site  has two protocols, one using sucrose  and the
other percoll?    The current recommendation is cresol red and reaction
mixtures are given on the web site.  At any rate this method has been used by
thousands of users of the Idaho Rapid Cycler with no complaints.  I know of
sevearal researchers who have used this on microtube cyclers such as Perkin
Elmer and Biometra.

web site is: http://128.110.195.115  or you can use any search program for
rapid cyclist.

Walt Schick



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