HELP:mRNA->cDNA->PCR

Robert Horton horton at molbio.cbs.umn.edu
Thu Feb 25 12:28:45 EST 1993


Dear Art again;

All that and I forgot to toot our horn about DNA compatible loading dyes!
This is a trick that helps a lot in analysing large numbers of PCR samples;
add a loading dye consisting of 60% sucrose/1mM cresol red to the PCRs
before running. Use ~2ul per 10ul reaction. Since you can add this to the
master mix before you do the PCR, it saves you the manipulations required
to add it independantly to your huge number of samples. Also, we routinely
color our cDNA templates by using 1% of the final volume of Schilling
yellow food coloring (mostly FD&C yellow #5, aka tartrazine). This makes
it easier to keep track of whether you've added template.

REFERENCE:
Hoppe et al., Gel loading dyes compatible with PCR. BioTechniques 12(5):
679-680 (1992).


- Bob Horton
(612) 624-3790



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