How can you troubleshoot issues with gel electrophoresis?

If facing poor resolution in gel electrophoresis, review the gel concentration and running conditions. Adjust parameters such as buffer concentration, voltage, and run time. Ensure the gel is properly cast, and consider using a higher percentage gel for better separation.

Regarding DNA, either digested or PCR products I always find pouring your own gel yields better and more size accurate bands, commercial ready to run gels are OK for fast results, but bands might be slurry and size might be off a little, for the latter always adding water to each well improves the quality, but still doesn't have the control that a "homemade" gel has, explanations for this is the power control and size and agarose concentration. Another experience or old school preference of mine it's the fact of being able to load larger DNA amounts in a self-prepared gel where one can optimize the well size to accommodate larger volumes, plus easier to excise DNA bands for further manipulations like gel extraction.

Uneven or distorted bands may result from uneven loading, improper sample preparation, or issues with the gel matrix. Verify that samples are loaded uniformly, and ensure equal concentrations. Check for air bubbles in the gel or anomalies in the casting process.

When bands are absent or weak, assess the integrity of the DNA or protein samples. Confirm sample purity, concentration, and preparation. Examine the electrophoresis apparatus for proper setup, ensuring electrodes, buffers, and power supply are functioning correctly.

When no bands are present on an electrophoresis gel, one of my initial troubleshooting steps is verifying the gel box has voltage flowing between the poles. The electrode wires can snap over time from consistent use or from mishandling. Additionally, I check for weak connections at the terminals as sometimes the electrode wires do not make good contact or are inhibited by corrosion. The orientation of the wires is also critical as reversing polarity will run the sample out of the gel and into the buffer. Lastly, the gel may have been run for too long or at too high of voltage and may have simply run off the gel itself. It is essential the electrophoresis conditions are monitored and adjusted to account for sample size and type.

Artifacts or background noise can arise from contaminants or improper handling. Employ high-quality reagents, ensure clean equipment, and handle samples with care. Evaluate the gel running conditions and consider optimizing staining and destaining procedures.

1?? Verify that the power supply is functioning properly. 2?? Check for buffer depletion during the run and replace if necessary. 3?? Ensure the gel apparatus is free from leaks. 4?? Investigate the condition of electrodes and buffer chambers. 5?? Consider using molecular weight markers to validate gel performance.