I got 60 micro-litre samples from IP and want to load them into single wells respectively. The problem is that the well can only hold about 30 micro-litre. How can I load 60 micro-litre sample to a single well in SDS-PAGE gel?

sir,
i'm not sure about what i'm saying but i think it may be possible to load 60
micro-litre sample by increasing the the thickness of the gel.sir,once again i'm clarifying that i'm not sure about it.

srini.

Hi,

you can tried to load your sample on a 'big gel' (about 7'*7'). The comb of these gel allowed to load easily 60uL.

Also, you can tried to lyophilied or dialysed/lyophilied your sample. With these manip, you can probably reduced the volume of your sample to 30 ul.

good luck

What I've done in the past (and it seems to work ) is to load 30 uL and then run the sample towards the botton of the stacking gel and load another 30 uL before it reaches the seperating gel. You have to be 'on-the-ball'; but it's always worked for me.

hi
if you use a small pa gel you can make a "double well" using adhesive between two teeth of the comb.

I would add your denaturing buffer to your 60ul of sample then put it at 99 degrees with the lid open - this will allow the water to evaporate and thus concentrate your sample.

Nay on Mar 31 2005, 03:13 AM said: