storing the loaded gel before running it - (Sep/03/2012 )
I have loaded my samples in the gels and all ready to go but the blotter machine did not work. I am wondering if I can store the gel that is already loaded with my samles and is in the running buffer right now? I do not think it is possible but I have a 26 lane gel and I really want to store it if I could and barrow a blotter from other lab tomorrow?
thanks
Just to clarify. Which machine isn't working? - SDS PAGE gel electrophoresis machine? or the Western transfer machine?
But I wouldn't recommend keeping the gel with samples loaded on it since the sample will diffuse in the gel - will either diffuse sideways or gets pulled down by gravity. So, best way is to reload another gel when you've gotten a machine that works.
It is the SDS PAGE gel electrophoresis machine. Thanks!
fhm on Tue Sep 4 00:42:39 2012 said:
It is the SDS PAGE gel electrophoresis machine. Thanks!
Sample will most probably start diffusing through the gel. Worst case scenario is you'll have merging bands or feint bands. I would recommend you restart the SDS with new samples. Did you prepare extra (backup) samples? This is always a good idea.
I totally agree with science noob. Moreover, I also do not support the common practice of storing SDS-PAGE gels (the ones with stacking and resolving parts). The stacking and the resolving gels have a different pH for a certain reason. Storing them in the fridge, even overnight, makes the H+ diffuse in between the 2 parts leading to decrease in resolution. Just a small comment.
Andreea
If you had no other option and needed to use this gel, I would consider making a mild fix solution (5% phosphoric acid) and only store it overnight. You would still have a mild amount of diffusion and the efficiency of your electrotransfer from your gel to your WB membrane would be reduced, but it would still work.
I store gels in ddWater overnight quite frequently and have never had any trouble with electrophoresis the next day, the stacking gel I use has the same pH as the resolving gel.
thank you guys. I really appreciate it.
proteaMatt on Tue Sep 4 20:19:45 2012 said:
If you had no other option and needed to use this gel, I would consider making a mild fix solution (5% phosphoric acid) and only store it overnight. You would still have a mild amount of diffusion and the efficiency of your electrotransfer from your gel to your WB membrane would be reduced, but it would still work.
this would be okay if the gel had been run. i believe that fhm said that s/he loaded the gel but couldn't run it.