3 replies to this topic

[Danny Chow]

I tried 150V and run the whole thing in ICE... but still ... smiling effect...

would it be the temperature? Should i not running the whole thing in ice??
cause most of the ... i mean all of the protocols didnt suggest me to run it at low temperature....

except buying a commercial gel, is there any other ways to solve the problem ?

[Bomber]

I haven't followed your previous discussions..., anyway I got rid of smiling effects by casting thicker gels (e.g 1.5mm)
Whenever I casted a thinner one I got the smilies.
Maybe worth a try.
Good luck.
I don't think it is the temperature/voltage... .

[mikew]

How to get rid of smiling gel.

#1. I assume you are using a BioRad system right?
#2. Biorad is conducive to "smiley gels". This is normally due to A) not all lanes being loaded
The ends of the gels leaked during casting C) Buffer is leaking.

If ALL of the lanes form a smiley face then it is most likely C.
This problem can be alleviated by filling the tank to the TOP. Not just the inside reservoir , the the whole tank.
This will inhibit the buffer from leaking.
Usually the 2 outermost lanes in BioRAD aren't so good due to some leaking of the gel into the spacer region.

[mdfenko]

Danny Chow, on Mar 30 2009, 04:33 AM, said:

I tried 150V and run the whole thing in ICE... but still ... smiling effect...

would it be the temperature? Should i not running the whole thing in ice??
cause most of the ... i mean all of the protocols didnt suggest me to run it at low temperature....

except buying a commercial gel, is there any other ways to solve the problem ?

150V is too much. run at less than 100V (cooling will help). this should eliminate (or, at least, reduce) smiling. it will take a little longer to run.

whole gel width smiling is caused by uneven heating of the gel (the center is warmer than the sides). the way to eliminate it is to ensure even heat distribution by limiting heating or by cooling.