Dear Lee

Which is the acrilammide% of this gel?

which running buffer did you used?

did you prepared the gel by yourself or it is a commercial gel ?

Manuele

Dear Manuele

i use 15% acrylamide gel

running buffer : 25mM Tris(pH8.3), 250mM glycine, 0.1% SDS

(10x stock: Trisbase 30.4g, glycine 144g, SDS 10g --> use after dilution to 1x)

i made gel , and use it within a 2weeks

gel prepartion

Stock

4X stacking gel buffer (500ml) 0.5M Tris-HCl (pH6.8) : 30.29g , 0.4% SDS 2g

4X running gel buffer (500ml) 1.5M Tris-HCl(pH8.8) : 90.87g, 0.4% SDS : 2g

Stacking gel 6ml

- H2O 3.63ml

- 40% Acrylamide 0.75ml

- Stacking gel buffer 1.5ml

- 10% APS 60ul

- TEMED 6ul

Running gel 25ml

- H2O 8.875ml

- 40% Acrylamide 9.375ml

- 4X running gel buffer 6.25ml

- 10% APS 250ul

- TEMED 10ul

thanks for attention

hi

1- run a samples on SDS page 15 %

2- check pH & composition of running buffer

3- run samples with lower voltage ( about 80 V )

4- put tank on the ice tray

good luck

Dear Haemin

did you normally store the gel for so long time?

Because when i did the gel by myself with a recipe similar to yours, i used it with in 24h.

What it is strange is also that the marker seems to run better than the samples. Are you sure that hte problem is not is the sample preparation?

Which loading buffer did you used? the samples were loaded well or are dense and difficult to be loaded?

Manuele