Gel in the attached image: 1% agarose, 100 V, 50 minutes, 100 bp ladder, 6ul was loaded in each well (1ul of 6X loading dye and 5 ul DNA).
I was expecting to not see any bands in this gel, as I'm testing a primer set that should not amplify the DNA in these samples (control).
Why am I seeing these streaks, starting from the well and then fading out? Is this something to do with the gel or the samples?
I had previously tried this (same conditions), but had loaded 10ul in each well (2ul of 6X loading dye and 8ul DNA, based on a colleagues protocol), and saw this pattern of streaking. I thought that maybe I was overloading the wells and that was causing the streaks. After fixing the dye:DNA ratio and lowering the volume loaded in the wells, I still see the streaking issue (in this image).
Any advice on where to start making modifications to my protocol or reasons why this may be happening would be greatly appreciated.
it looks like high molecular weight dna. You may be amplifying too much dna.Try running an equivalent amount of the streaky dna sample that has not been amplified and see if you get the same effect
1) The concentration of the DNA appears to be higher.
2) The smeared bands indicates the partial DNA degradation.
So better check the DNA concentration and 260/280 ratio before the next experiment. Good luck Christina Desnoyers
It may be due to high concentration of DNA. dilute the concetration and adjust the proper concentration using nanodrope
Good luck
Degradation in DNA. 1% gel is too low with such a high voltage. I think you try 3% conc. With 65 V for 120 minutes
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