The quality of my gel has been progressively getting worse. I am only attempting to check the quality of the DNA, so even in an example such as this, I can still confirm the topmost band that I need within most of the wells. However, the amount of smearing I am seeing downstream has been becoming more prevalent, and I am unsure if this has something to do with the extraction method or in the preparation of the gel. The gel is 1% agarose, 10,000X dilution of SYBR Safe DNA Gel Stain, 1X TBE buffer, 5 uL of 1 KB ladder (leftmost), and 6 uL of varying quantities of DNA (~5-100 ng/mL, depending on how well the extraction went). Is there a problem with the gel, something to do with the extraction, or some other issue?
Considering that even the ladder is not resolved, I would say the problem is probably with your gel. In one front, check the quality of the agarose and the TBE buffer (the latter can sometimes present mold if not made fresh). On the other, monitor the electric current during the run; high currents can excessively heat the buffer, causing the smearing. Reduce the voltage if necessary.
Also check this thread:
https://www.researchgate.net/post/Does-anyone-have-an-experience-on-how-to-overcome-blurry-fuzzy-or-smeared-bands-sudden-change-in-quality-of-gels
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA