6x Loading Buffers

Robert Horton horton at biosci.cbs.umn.edu
Fri Oct 13 20:34:39 EST 1995


Frank P. Johnston (fpjohn at msn.fullfeed.com) wrote:

: If anyone has any "recipes" for different buffers could you please send 
: them my way? Oh yeah, the company we ordered the primers from doesn't 
: sell the buffers in their catalog...


Have you tried our PCR-compatible loading dye? Dissolve ~1mM cresol red
(free acid, available from Sigma or Aldrich - this is near its
solubility limit) in 60% sucrose. You can add it to your PCRs before
cycling, so you can put it in the master mix and you don't have to add
dye to each sample afterwards. See:

Hoppe, B.L., Conti-Tronconi, B.M. and Horton, R.M. "Gel-loading dyes
compatible with PCR". BioTechniques, 12(5):679-80, 1992.

Cresol red is a pH indicator. It is red above about pH8 (as in regular
PCR buffer or TBE), and yellow below. It migrates at somewhere around
300bp in a 1% agarose gel.



Robert M. Horton (PhD!) /\ "Crash programs fail because of the theory that
U of M Dermatology Dept || with nine women pregnant you get a baby a month"
Box 98 UMHC, 4-154 PWB /||\ -W. von Braun.   Disclaimer: "Bob who?"
Minneapolis, MN 55455   ^^   horton at biosci.cbs.umn.edu       (612)625-8941







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