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bands bend in SDS PAGE - (Sep/27/2006 )

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hi all
recently i noticed that my bands specially the first and second lane bend to the right after electrophoresis before it was straight coulmn if u can say but now it tilts at the lower end of the gel?
why?
thankx in advance

-spanishflower-

Aha, smiling gels. This is nothing new. I routinely run 6-10% vertical denaturing polyacrylamide gel at temperatures between 50-60 deg C.

There is normally always some 'smile' at the base of the gel or at the very edges. This is normally due to non-uniform heat distribution through the gel matrix. The edges will be slightly cooler and therefore have a higher resistance compared the centre of the gel. This means that the lanes at the sides migrate ever so slightly slower.

-Doc_Martin-

thankx doc martin
but i think i didnt make myself clear because of my english
i know the smiling gel thing but my problem isnot that
i am running a prestained protein marker and after the 50kda band the bands started to go to the right
then the next lane i run some bacterial lysate and after this band also it tilt to the left????
my professor has no answer so please can u help???

-spanishflower-

May be the resolving gel was not mix properly before pouring.

-Minnie Mouse-

thankx minnie
i have some ideas in my mind i will check
1-as u said about mixing the gel well
2- remake 10x running buffer
3- mix well 1x running buffer
4- put running buffer in the outer chamber withinn 1 cm from the gel plate
5- apply 1x running buffer to the empty well in the gel
i will let u know the results.

-spanishflower-

can it be the problem of the comb? unsure.gif ....all I can think of other than that is non-mixed gel as Minnie said.....not too much help sorry.

-Kathy-

it happened to me, it was where I put the pipet to pour the gel.
now I pour exactly in the corner.

-Missele-

QUOTE
it happened to me, it was where I put the pipet to pour the gel.
now I pour exactly in the corner.


i pour directly from the tube not using the pipet, but first from the corner then all throught the glass from the top.

-Kathy-

Only to your right hand side? not on both sides?

-genehunter-1-

and finally a silly remark,

if this is a persistent problem, independent on which corner the gel is poured (which is most likely the cause), you might need to look at the electrode of the gel tank.

Is that little bit of wire still okay, and not snapped/shortened.. If the wire is damaged, the electric field will not be uniform.

-perneseblue-

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