Adding loading/tracking dye to PCR mix? - (Jul/10/2011 )

Hi guys,

I was wondering, can normal 6x tracking dye be added to PCR mix before going for thermal cycling conditions?

What is the reason for doing that? The loading dying may interfere your PCR reaction.

one of my boss is doing on PCR chip. so he wanted me to add some dye for the ease of viewing.

and yes, loading dye interferes the PCR. just saw the PCR result just now. was using loading dye from Fermentas.

any solution for that?

You can buy polymerase that has loading dye included in buffer, like GoTaq Green from Promega.
Or try adding only the dye (bromphenol blue or xylene cyanol) to to mix, not loading buffer (which contains glycerol or something like that in addition), as I understand you only need the mix to be colored, not dense.

oh yes, Sigma's RedTaq has loading dye added and is very good.

I use Eurx Perpetual Taq polymerase (hotstart), with supplied coloured buffer. Works very well for me, and much more cheaper than Promega GoTaq.

Adrian K on Mon Jul 11 17:31:48 2011 said:

That was just an example. I don't use it. Sure there are others that may be better and cheaper (the performance of GoTaq really didn't impress me).

Trof on Tue Jul 12 08:58:29 2011 said:

Agree with Trof. Thats why I switch brand

disagree with you, gentlemen...
GoTaq Green Mix -Promega, works very fine with me ...

p.s : i felt that i must add my comment, for Promega's ...
since, it has been serving me!

i never thought of adding just dye alone to the PCR mix

thx guys