hi

you can do :

1- Let agarose solution cool down until it has completely solidified.

2- check TAE have right composition (1X)

3- don't change the voltage during the gel ruining procedure (80 V is ok) .

4- check the mA of POWER instrument during gel ruining , it must be constant (not variable) .

good luck

you'd better boil the agarose fully. I guess the agarose was not getting solid well.

hi

you can do :

1- Let agarose solution cool down until it has completely solidified.

2- check TAE have right composition (1X)

3- don't change the voltage during the gel ruining procedure (80 V is ok) .

4- check the mA of POWER instrument during gel ruining , it must be constant (not variable) .

good luck

Thank you very much, Ma Dejun and Reza Hadavi for your time and kindness. However, the agarose has cooled down and completely solidified!!!. Moreover, I noticed that the buffer is heating up and run is very very slow.

As the buffer is getting heat，it is notable to change the electrophorsis instrument or perform the agarose in ice. beacuse the heat could locally melt the agarose and then curve the band.

Make sure you have enough buffer in your tank. You should have almost 1 cm of depth above your gel otherwise the resistance will be quite high and you will heat your gel.

Run your sample slow (90V) until it enters the lane from the well, then increase the voltage to 110-120V.

I would also recommend changing out your buffer for fresh buffer and checking to make sure there is no deposition on your gel box wires.

Dear Ma Dejun and Jordan Yaron for generosity and kindness. When I used EB instead of safe stain, I get good results.

I never thought that the dye could have such a great effect on the band. maybe I

always used EB. haha

Are you running the samples already stained? If so, run them unstained and stain your gel after the fact. I have run into that issue before, and post-staining usually corrects it.