I’m currently conducting research on telomeres in THP-1 cells. During my gel electrophoresis experiments using a 1% gel, I’ve encountered an issue where the DNA seems to be stuck at the top of the well. Additionally, there are two unexplained lines appearing within the DNA smear. Could anyone provide insights into what might be causing these issues?
Your samples contain proteins or other compounds coating the DNA and preventing its migration.
You need to provide more information. Are these digested with any restriction enzymes, if so which ones? This almost looks like undigested genomic DNA, in which case it will never enter a 1% agarose gel. The smears on the sides of the wells are just an artifact of running large DNAs on a gel designed for fragments in the range of 100-10,000 bp or so.
The issue seems to be in your DNA sample. Please check the purity of your DNA samples and run only if the ratio of 230/280 value is 1.8.
I am agree with all who answered to your question you must check the purity of DNA samples and all other things and you focus that you work on the Teleomers and this region is very important region to DNA as follows:
Telomere. A chromosome is essentially a long, long piece of DNA that has really wrapped up and compacted on itself until it looks like the structure you probably picture when I say chromosome. The problem is that the long piece of DNA has two ends, and they're just hanging out there loose. A lot of things could go wrong if those ends aren't protected. They could get cut off, or they could join onto other loose DNA ends, which would be a problem for the cell.
I would advise you check the purity of your sample as there might be some impurities or aggregates present and with the smear I reckon there might be some degradation of your sample and if you used any restriction enzyme try to optimize it. wishing you all the best in your paper
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