Gap between bands too small - DNA Gel Electrophoresis (Oct/31/2007 )
I ran a DNA Electrophoresis with Wild Type and KO Mouse DNA samples on 2% Agarose Gel. Everything was fine but the bands are too close together. Wild is somewhere around 170 and KO is just a little lower; so much that it is confusing to say they are different.
Is there any way I can make the band separate further?
I thought increasing the time for electrophoresis will help. Should I also change the concentration of Agarose?
Hi, try to use more concentrated 3 % agarose gel. You should be able to distinguish 10 bp difference in length. Other possibility is PAGE.
2-3% metaphor agarose probably will do the job. Can you cut one of the fragments differently from the other? Is there a region which will bind an oligo differently?
Make 3-4% agarose gel, this should help to differentiate the bands.
Thank U for ur replies.
I shall attempt increasing the concentration of the gel and tell U how it goes. But, allow me some time before I get another DNA sample.
Also running electroforesis on lower voltage (0,7 V per cm of electrode distance) can increase the resolution of fragments (and running it long enough since the bands would be more blurred), but in this case trying more concentrating agarose is the first thing to do.
Thank u Trof. I will try that.