I am using Bio-Rad StainFree AnykD precast gels, and in many of them (I'd say four out of six of the box I have run) the dye has migrated weirdly - instead of being in a nice blue hazy band it looks as though there are two blue bands - one in the gel, and on almost in front of the gel (but behind the plastic). This also happened to another lab member once who was using a different box of gels, different apparatus and buffers.
I feel like it might be the speed of the run? 200V for 30 min. We use Tris Glycine buffer. Has this happened to anyone before? Any idea of what causes it?
Hi,
I would bet on pH issues too. You won't have many choices but to test again your buffers.
Best of lucks.
Agree with the above. Have also run these and other pre cast held at 200v for 30 min and that should be fine in general but a word of caution: running held at high power quickly will exacerbate the sorts of problems you are describing especially when at near or beyond expiry date
Along with above suggestions i will suggest you to reduce your sample loading time. If u r taking more time for sample loading than usual i feel the bands will get diffused and may not have proper dye front. I will also suggest to reduce the power and increase the run time.
Thank you all, this has been very helpful. The gel pack was not expired but not too far from it - in the past we have reused buffer no problems but I did test and found that my pH was a bit out and even further out when the buffer was reused. I have not noticed the problem again with my properly pH'd buffer, but I am also using a new packet of gels.
I also think it is wonderful that I can get help from people with such diverse backgrounds and experience levels on ResearchGate. Thank you for your contributions.
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