gel electrophoresis - problem with stacking (Jan/18/2007 )
QUOTE (mdfenko @ Feb 1 2007, 07:50 PM)
QUOTE (louise00 @ Feb 1 2007, 11:01 AM)
I see. This time i did have them in the fume hood actually because someone said i should leave them in there beacuse of the acrylamide. Do you think that could have been it?
yes, it may be. with the air currents in the hood you may be getting uneven cooling of your gel, resulting in uneven polymerization. you would have to allow the gel to polymerize for extended periods of time (until it is complete).
there is no need to polymerize in the hood. once acrylamide is in solution there is no danger of fumes or airborne particles (unless you sneeze into the tube, flask or plates) from the acrylamide.
Oh right, i didnt realise that.
well I have to make two more tomorrow so i will mix well and keep them out the fume hood!
Thanks very much for your help, its been quite maddening!
-louise00-
QUOTE (biotech! @ Jan 18 2007, 08:46 PM)
Hi, does anyone know why it can be happening this: When doing gels I have no problem with the polimeration of the gel from below but, the stacking keeps liquid or takes hours to be made. 
......it's really weird since I use exactly the same things that for the first one except for the buffer (pH 6.8 instead of 8.8).
Now I found a solution....I use the same pH 8.8 buffer for both parts and it works.......
I' don't know what might be happening, I'already checked the pH so that's not the reason.
By the way, do you know why the buffers have to have different pH??
well, hoping to find the reason!! ;)
......it's really weird since I use exactly the same things that for the first one except for the buffer (pH 6.8 instead of 8.8).Now I found a solution....I use the same pH 8.8 buffer for both parts and it works.......
I' don't know what might be happening, I'already checked the pH so that's not the reason.
By the way, do you know why the buffers have to have different pH??
well, hoping to find the reason!! ;)
don't use the Same PH 8.8 buffer for the stacking , make sure that you made the 10% APS fresh in distilled water , you can increase the TEMED which will solve your problem , or alternatively you can use 5% stacking gel,
-Amit nayak-
Use only fresh APS and mix well the mixture
QUOTE (biotech! @ Jan 18 2007, 04:16 PM)
Hi, does anyone know why it can be happening this: When doing gels I have no problem with the polimeration of the gel from below but, the stacking keeps liquid or takes hours to be made. ......it's really weird since I use exactly the same things that for the first one except for the buffer (pH 6.8 instead of 8.8).
Now I found a solution....I use the same pH 8.8 buffer for both parts and it works.......
I' don't know what might be happening, I'already checked the pH so that's not the reason.
By the way, do you know why the buffers have to have different pH??
well, hoping to find the reason!! ;)
......it's really weird since I use exactly the same things that for the first one except for the buffer (pH 6.8 instead of 8.8).Now I found a solution....I use the same pH 8.8 buffer for both parts and it works.......
I' don't know what might be happening, I'already checked the pH so that's not the reason.
By the way, do you know why the buffers have to have different pH??
well, hoping to find the reason!! ;)
-biomolman-