Posted by
Coutu, Cathy on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Problems-with-Red-Safe-tp6927958p6929551.html
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Hi Cláudia,
We use EnviroSafe Gel Dye (Helixx Technologies cat #HDS001-3 for international orders, $175/3 ml, use 5 ul/30 ml agarose gel).
We run about 320 ng of digested DNA in 7 ul in a 2mm wide x 1 mm thick well.
We can clearly see the bands (usually looking for about 4 bands).
If you are looking for many bands, each band will appear less bright and you'll need more concentrated DNA.
If you are using a larger well, each band will appear less bright and you'll need more DNA.
You didn't mention if you incorporate the dye into your gels prior to running them, but EnviroSafe was the best dye we found for this purpose.
Cathy
Cathy Coutu, M. Sc.
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-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Claudia Campos
Sent: October-25-11 1:59 AM
To:
[hidden email]
Subject: Problems with Red Safe
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Dear all,
I know this is a bit out of the list scope, but I have to share it.
Our institute shifted from EtBromide to Red safe about a year ago. After
this, we had to create a specific room for Et Br because nobody could do
regular cloning with Red safe. Not so good, but feasable.
In our lab we also realized that whenever you do a restriction, just to
check minipreps, you ALWAYS have to clean up your restriction with a column,
otherwise you don't see any DNA on a Red safe gel (and it really looks like
an empty gel!).
The bad thing is that when i got in contact with the company, they just
replied that when testing the product they always purified DNA, and that we
should concentrate it. They didn't provide any further explanation on why
the gel seems completely empty if we don't clean up our restriction (an
extra step that costs money and time!).
Does any one else has the same problem?
If it goes on like this, we may seriously consider shifting back to Et
Bromide and letting go red safe...
Thanks for any feedback!
Cheers
--
Cláudia Campos
Lab manager - Cell Biophysics and Development Lab
Instituto Gulbenkian Ciência
R. Quinta Grande, nº6
2781-154 Oeiras PT
Tel:+ 351 214 464 608
http://www.igc.gulbenkian.pt/research/unit/38