My DNA shoots out of the wells in my Agarose Gel... why? - Agarose Gel Loading Help (Jan/29/2007 )
Gel is ~ 7mm thick, wells ~ 5mm deep, 5mm wide, 1mm thick, 0,8%, 0,5X TBE, ~10-20 ul samples. I will try again with more drying. And pippet more slowly.
I've tried "semi-dry" electrophoresis totay, without submerging wells but gel was in contact with buffer . It... works. And my menthor said that's OK. But I'm still not happy *perfectionist*.
BTW how much DNA is usually lost when precipitating with EtOH? M.
I would retract my earlier post...I don't even see how it would work if the gel weren't submerged
The gel is 3/4 submerged when loading. It worked. After few minutes of electrophoresis, when samples entered the gel I've added some more buffer and covered is completly. M.
Not meaning to insult your intelligence, or ability to read a protocol, but are you using DNA loading buffer? Protein loading buffer looks pretty similar, but I wouldn't want to mix them up.
Can you get loading buffer from another lab? Is anyone else in your group having troubles?
Yes, I'm using the right buffer. No proteins in our lab, just DNA and RNA. And we are using home made buffer. It worked fine until now. Sometimes others are also having problems, but they seem to occur randomly. I will try the same thing again monday... M.