Ethidium Bromide vs Sybr Green stain (normal PCR) - (Nov/01/2005 )
Our lab is trying to minimise the amount of ethidium in use. Our standard method at the moment is to add the ethidium to the gel (and the buffer if a long run is required). The problem with this is that we have a lot of students come through and their technique is sloppy (regardless of numerous reminders).
I would prefer not to post-stain the gels as they can't be run longer. I am aware that some people add the Eth Br to the loading dye and run it that way. Has anyone any comments on this technique regarding the concentration of the Eth Br used and the resolution of the bands?
The other option I'm eager to learn more about is using Sybr green. The protocols I have found so far recommend post-staining which brings up the problem of not being able to run the gel further. Has anyone tried using Sybr green in the loading dye? I know that it's detection can be tricky depending on the type of gel doc system in use.
I understand that the danger of using Eth Br is contentious, but better safe than sorry
Why can't you run the gel longer after it is stained, whether EtBr or sybr green?
We routinely add EtBr to the gel (but not the buffer) then run it and if we want to run farther, or if the EtBr migration is above the bands so they "disappear" then we just re-stain the gel with EtBr...
i do this on my RNA loading procedure in agarose formaldéhyde gels. I add 1µl of 0.4mg/ml BET solution to20µl sample.
[font=Garamond]We use SYBR Green for our gels all the time. We normally add it straight to our loading buffer or we post-stain using it. It works really well and it's much more safe for students to use. You also don't have to worry about the waste.
SYBR Green is comparatively expensive but works really great after staining the gel . Otherwise we use EtBr added to gel.
hi to everyone
does anyone know if SYBR green stains protein becides nucleotides?