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Adding loading/tracking dye to PCR mix?


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10 replies to this topic

#1 badguy

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Posted 10 July 2011 - 06:36 PM

Hi guys,

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



#2 pcrman

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Posted 10 July 2011 - 10:06 PM

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

#3 badguy

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Posted 10 July 2011 - 10:30 PM

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?



#4 Trof

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Posted 11 July 2011 - 06:34 AM

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.

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#5 pcrman

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Posted 11 July 2011 - 07:12 AM

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

#6 Adrian K

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Posted 11 July 2011 - 09:31 AM

I use Eurx Perpetual Taq polymerase (hotstart), with supplied coloured buffer. Works very well for me, and much more cheaper than Promega GoTaq.
:blink:
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

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#7 Trof

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Posted 12 July 2011 - 12:58 AM

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

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).

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon


#8 Adrian K

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Posted 12 July 2011 - 01:06 AM


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

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).

Agree with Trof. Thats why I switch brand ^_^ :lol:
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434

#9 nightingale

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Posted 14 July 2011 - 11:40 AM

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!

Edited by nightingale, 14 July 2011 - 11:42 AM.

" The more you learn, the more you realize how little you know ... "

#10 badguy

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Posted 14 July 2011 - 07:27 PM

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

thx guys



#11 klinmed

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Posted 17 July 2011 - 01:38 PM

Add some cresol red dye to your pcr mix. This stain migrates at 1000bp on 1% agarose gels and does not inhibit taq. If you need a little more density for easy gel loading add 5% glycerol also.

See also: (1)  Hoppe, B.L. et al. (1992) Biotechniques 12, 679-680
Gel-Loading Dyes Compatible with PCR.

Hope this helps




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