13 replies to this topic

[ros]

Hi all,

I've just started in a new lab, and here they re-use their 1X TAE for DNA gels up to 5 times. I was under the impression that it should only be used once, however  TBE could be used around 3 times.

So, should you, or should you not re-use TAE?

Thanks alot....

Ros[B][COLOR=red]

[labrat]

I guess we use at least 10 times before someone changes the buffer.  I think it is OK to continue use it if DNA can be well separated and bands are not distorted.

[ros]

hmm...interesting. thanks. 10 times - wow!!

i just used google to do a search, and found the commercially available 50XTAE from eppendorf should only be used once. presumably its the same recipe as standard TAE...

Website:
www.eppendorf.com/script/cms-newspic. php?id=1771&inline=1&col=DOWNLOADFILE

quote:
"For agarose gel electrophoresis, 50x TAE should be diluted to a working concentration of 1x. The TAE buffer should be
replaced after each electrophoresis run due to the anode solution becoming alkaline and the cathode solution acidic, which
results in lowered DNA mobility."

but if it works, then may as well.

thanks again...

[labrat]

Quote

"For agarose gel electrophoresis, 50x TAE should be diluted to a working concentration of 1x. The TAE buffer should be
replaced after each electrophoresis run due to the anode solution becoming alkaline and the cathode solution acidic, which
results in lowered DNA mobility."

So you will buy more from them, they will make more money

[zhgljj1998]

ros, on Jan 27 2005, 10:44 PM, said:

Hi all,

I've just started in a new lab, and here they re-use their 1X TAE for DNA gels up to 5 times. I was under the impression that it should only be used once, however  TBE could be used around 3 times.

So, should you, or should you not re-use TAE?

Thanks alot....

Ros[B][COLOR=red]

if you do not cut your gel, then reuse it until someone else change the buffer.

[Al Bunsy]

Quote

anode solution becoming alkaline and the cathode solution acidic

Heh... It looks funny. No one can realize that to make them the same pH one hase just to mix them.

[george@CASE]

I use 0.5X TBE to run my gels and I only use it twice.

Aside from remixing them to stabilize pH, one has to take into account that the tank buffer gets more concentrated the more you use it.

The little bubbles that come up from electrolysis of water.

[jy2004]

It is OK for us to re-use 0.5X TAE for more than 10 times in our lab. But we don't keep the gel in the gel box after running gel each time and take the gel out, cut the dye band out. The rest part you can reuse.

[methylnick]

Hi all, in our lab we re-use our homemade buffer until it runs out of "kick" that is to say the current exceeds 200mA when set at 120V.

I have worked in a lab where the gel itself is remelted and reused!!

Nick

[chandima]

hi
in our lab we re-use the home made buffer and the gells as much as we can for routine work. in fact i pour my gells on glass plates to make them thin and re-use it unless it is not broken in handling.
chandima

[fred_33]

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)

Edited by fred_33, 21 February 2005 - 02:28 AM.

[expresson_help]

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!

[ros]

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!

[Beautiful_Tits]

labrat, on Jan 27 2005, 11:04 PM, said:

I guess we use at least 10 times before someone changes the buffer.  I think it is OK to continue use it if DNA can be well separated and bands are not distorted.

yes, i think this will due