inert dye for real-time pcr

[jcww99]

I'm looking for an inert dye to add to my colorless real-time pcr
mixes to be able to check the performance of a pipetting robot. Of
course there are commercial solutions like Thermo's Absolute Blue but
these are not an option. Years ago in this group tatrazine was
mentioned as a possible dye. Does anyone know more ????

Thanks to you all

Emilio

[Tom Knight]

Cresol Red is another option. Just a quick warning for a problem we
encountered, which is that some of these dyes fluoresce. This is not
normally a problem, but can completely trash a sequencing reaction if
dye manages to get into the DNA template. There may be issues with
real time PCR detection as well.

[Tim Fitzwater]

"I'm looking for an inert dye to add to my colorless real-time pcr
mixes to be able to check the performance of a pipetting robot."

There are published reports of two different red dyes used for this
purpose.

1) 0.01% Phenol red (R. J. Klebe, S. A. Rodriguez, M. L. VerBeek and T.
A.

Giambernardi, 1999. Simple method for "hot-start" RT-PCR.

BioTechn. 27(6):1108-1110.)

2) Carl Wittwer's laboratory uses 0.1 mM Cresol Red for the same purpose

(The Rapid Cyclist, Spring, 1994, page 9).

Tim

[peter]

Since PCR mix is H2O based you can try any number of food color dyes,
provided that they wont interfere with the emission/excitation spectra
of your real-time detection system.
Peter