Reuse TAE buffer and agarose gel? - yes or no... (Jan/27/2005 )
QUOTE
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?
The thing is this one... EtBr runs in the direction opposite of dna. So if you rerun the gel later, much of the etbr has migrated out of that area and cant bind to the dna. That is why when we cast a gel with two rows of lanes, we run the lower lanes first, then the upper.QUOTE
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.
Our lab stock of Etbr is 10mg/ml, we use 5ul in a 100ml gel... so I guess your etbr is just quite dilute. I don't remember what Sambrook recommends.
-pBluescript-
Yeah one can use TAE buffer more than ten times for visulalization purposes, as Nick ve mentioned I ve seen people melting the gel and reusing that too..
In our experiments sometimes we can't follow the manual or the other guides exactly word by word, they ve mentioned that to promote the sales of their products.
Julianne, I guess you got some answers from the members for your query "How to cut cost in a molecular biolog lab?" through this question too.
have a nice time
smiles
Saint
-Biosaint-
I also tried reusing agarose gel by melting used gel. It works fine although it is not necessary for me. A concern I have is if the gel has been stained with EB, when it's heated, you may breathe EB into your body.
-pcrman-