remelting agarose gels with gel red

[spongya77]

I'd like to try it -my advisor does not want to buy too much agarose, but first, I'd like to ask for some advice from people who did it before.
Anything particular I should be looking out for? Or just melt the gel, and recast?
Thanks

[pigfarmer]

Originally Posted by spongya77

I'd like to try it -my advisor does not want to buy too much agarose, but first, I'd like to ask for some advice from people who did it before.
Anything particular I should be looking out for? Or just melt the gel, and recast?
Thanks

I always use My gels multiple times, as I only run maybe half a dozen samples at any given time. I have found that the signal with Ethidium Bromide is good to run a gel numerous times(usually 4-6 runs and I am out of room on my gel); however, the gel red signal is almost null after only running a gel once. I would assume you have to add new gel red every time you re-melt, but perhaps it would save more if you just used Ethidium bromide and re-used the gels you make multiple times?

[spongya77]

Thank you for answering.
Adding gel red is no problem; however I just tried it last week, and the DNA moved about 10 mm in two hours... (I have this aversion to EtBr -and so has everybody else working around the gel equipment...)
I guess something did not work out. Is it possible that so much buffer evaporated as I remelted, so that the gel became really concentrated?? Sounds very unlikely. Maybe it dried out in the fridge - in any way, how can I make sure that the gel's concentration is consistent after remelt?
Thank you

[talkingtree]

remelting gels? that's very interesting, how do you remelt gel to be reused? just microwave until it melts? but what happened to the dna that was in there?

[pigfarmer]

Originally Posted by spongya77

Thank you for answering.
Adding gel red is no problem; however I just tried it last week, and the DNA moved about 10 mm in two hours... (I have this aversion to EtBr -and so has everybody else working around the gel equipment...)
I guess something did not work out. Is it possible that so much buffer evaporated as I remelted, so that the gel became really concentrated?? Sounds very unlikely. Maybe it dried out in the fridge - in any way, how can I make sure that the gel's concentration is consistent after remelt?
Thank you

Seems that if so much of the buffer evaporated you would be able to see by eye that the volume was too low. How many times after remelting does this happen? When I used to remelt with EtBr I would only remelt once then make a new gel. When you save the gel to remelt you should also make sure not to leave it open, put in in an erlenmeier with parafilm over the top so it won't evaporate?

[jiajia1987]

Originally Posted by talkingtree

remelting gels? that's very interesting, how do you remelt gel to be reused? just microwave until it melts? but what happened to the dna that was in there?

Yes, I am pretty curious myself about this topic too. Can anyone answer to the questions above?

[pigfarmer]

Originally Posted by jiajia1987

Yes, I am pretty curious myself about this topic too. Can anyone answer to the questions above?

Its as said. Just re-melt the gel in the microwave, and add again EtBr or whatever you are using.

[jiajia1987]

So, if say... I have ran my samples in this particular 1% gel, and after using it (with all the loading dyes, DNA ladder and what not), I just melt it in the microwave again? The heat destroys the DNA ladder and loading dyes etc, and that's why the gel can be reused, right? I just need to chop the gel up into pieces and place them in a glass bottle and melt it? How about the buffer content lost? How many times can the gel be reused?

[spongya77]

Originally Posted by pigfarmer

Seems that if so much of the buffer evaporated you would be able to see by eye that the volume was too low. How many times after remelting does this happen? When I used to remelt with EtBr I would only remelt once then make a new gel. When you save the gel to remelt you should also make sure not to leave it open, put in in an erlenmeier with parafilm over the top so it won't evaporate?

It was the first time
I guess I'll just have to try it again. I appreciate your time

(To answer others' question: just break up the gel, so you can stuck it into an Erlenmeyer, and into the microwave. The DNA should not cause too much trouble -the background will be higher, and you probably should not use the gel for isolating and whatnot. Only for "dirty" applications.)

[jiajia1987]

Originally Posted by spongya77

It was the first time
I guess I'll just have to try it again. I appreciate your time

(To answer others' question: just break up the gel, so you can stuck it into an Erlenmeyer, and into the microwave. The DNA should not cause too much trouble -the background will be higher, and you probably should not use the gel for isolating and whatnot. Only for "dirty" applications.)

Thank you!