After the first (agarose or SDS-PAG) electrophoresis, if I run the same gel again with an-overtime so that everything in the gel will now in theory come off the gel (i.e. electro-rinsing), then could such gel be re-used for the next experiment?
If not, what would be the major limitation?
For agarose gel it is possible. Many people do it! For Acrylamide gel I don't think you can remove all proteins by running overtime. High molecular weight proteins don't move well in the gel, maybe they need days to be removed! Of course it depends also from gel concentration!
For agarose gel it is possible. Many people do it! For Acrylamide gel I don't think you can remove all proteins by running overtime. High molecular weight proteins don't move well in the gel, maybe they need days to be removed! Of course it depends also from gel concentration!
For SDS-PAGE, the problem is not a problem of protein but not of pH of buffer, it is a discontinue migration system, but after migration, all the gel have one pH and buffer, due to the migration of the electrolytes.
I've reused ethidium-stained agarose before for routine gels, but never with the same casting. We would just break up the gel, and put the pieces in a flask. Just before use we would re-melt it in the microwave and re-cast. Be careful of the steam, as it will contain ethidium. This has the disadvantage of causing a gradient in background because of the ethidium migrating in reverse of the DNA as David mentioned.
Theoretically , yes u can..depending on the type of gel and percenatge of gel, the protein would eventually come out and you could reuse it. But i am not sure how you would be sure as to whether there is any remnant protein bands at all left in the acrylamide gel..maybe after 2 days you could re run and see if you get any band at all. If not you could reuse it..but i am curious to know why you would need to reuse such PAGE gels, which could be brought as pre-cast gels or could be freshly cast within40 minutes ....
what about reuse the agarose gel when we used Saber Safe for Staining
Can agarose gels be cast with SYBR® Safe DNA gel stain in them?
Yes. Simply substitute a SYBR® Safe DNA gel stain solution for the buffer when preparing the molten agarose. If using the 10,000X SYBR® Safe DNA gel stain concentrate, dilute the concentrated stain 1:10,000 in agarose gel buffer (e.g., 1X TBE or 1X TAE) and add the buffer/stain solution to the powdered agarose. The agarose/SYBR® Safe DNA gel stain mixture may be heated briefly in the microwave.
The company strongly discourage the reuse of SYBR® Safe DNA gel stain, as this practice significantly lowers sensitivity.
Similar to ethidium bromide, SYBR® Safe DNA gel stain runs away from migrating DNA. This has no practical effect on the use of gels cast with SYBR® Safe DNA gel stain, as only the very bottom of the gel will have a lower concentration of stain. This effect can be partially counteracted by adding SYBR® Safe DNA gel stain to the running buffer
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