Reuse TAE buffer and agarose gel? - yes or no... (Jan/27/2005 )
hi
in our lab we use the buffer as far as 20 times or more. The buffer is changed when the colour has changed to blue due to the bromophenol blue used in loading buffer. But don't forget to remove ethidium bromide of the buffer (using columns from schleicher and schuell or activate carbon)
Eppendorf probably just want you to buy more of it!
I generally reuse gels until I run out of lanes - however, if they do go off, it isn't the resolution that goes, but for some reason the samples don't sink into the lanes as well after a while. Not sure if that's just me though!
so how do you reuse a gel? do you just run your samples off the bottom and then reload new ones? i take it you dont add ethidium bromide to the actual gel itself before it sets? how do you get rid of the ethidium anyway - whether you add it to the gel or whether you stain your gel afterwards?
i figured eppendorf just wants you to buy more buffer, but i make my own anyway - someone just told me once that you cant reuse it! evidently, they were very wrong!
yes, i think this will due
In our lab we re-use our agarose gels, and our buffer. The gels we simply re-melt in the microwave, and add a little buffer if they are getting too thin. The buffer (0.5X TBE) we re-use until it gets too blue, like someone said above. However, we only use the gels for visualization, of microsatellites mostly.
-Peace
P.S. We add the Ethidium Bromide directly to the gel after it has cooled down a bit but is still liquid. 400 ul on the first use, and 200 ul each additional use, at a [] of .01mg/ml if I remember correctly.
Hi,
We use our TAE many, many times until they make me (the student) change it. We also re-use our gels but only within a few hours of making it and i dont notice that it makes a difference to the quality.
You add 400ul of EtBr to your liquid gel mix? Are you making a huge amount of gel? We typically make 75ml of 1% Agarose gel (enough for 2 gels), and only add 1-2 ul of EtBr at 0.01mg/ml.
We routinely run several gels through 1x tae buffer. I keep an eye on the volume and change it if it has evaporated. Also, if I plan to cut a band out for cloning I use clean buffer. Works for us, at least... - Jen
Hi,
we use our TPE-buffer at least for 5-10 gels, depending on how much bromophenolblue
diffuses into it from the gel.
If the buffer is to "blue" it will quench the fluorescence of bands with a low
amount of DNA, which I frequently have during work with low copy
number plasmids.
We also stain our gels after electrophoresis and cut out the lanes with DNA.
Our "lab security guy" is quite concerned about the mutagenetic effects of
ethidiumbromide, so we discard the stained gel pices after use.
Bye,
Hennry
Oh, think my Lab has reused the buffer for at least 10 times. I just add some more TAE buffer to cover my gel.
Just wonder how many times can you reuse the gel ( as in the casted gel, not recasting the gel again)? My lab taught me to use the unused gel wells, however the effect isn't that good as first time. I presume that the effect of Et Br detoriates as time goes by, right? or by the no. of uses?