Smeared DNA Agarose Gel Electrophoresis Troubleshooting

Smeared DNA Agarose Gel Electrophoresis Troubleshooting 

{mosloadposition advert1} 

If your DNA has smeared on your agarose gel electrophoresis you may have the following problems:

• You loaded and ran degraded DNA. Make sure you run Avoid nuclease contamination.
• You loaded too much DNA into the gel well. Decrease the amount of DNA you are loading into each lane. Don't exceed more than 50ng/lane or band.
• Your samples had too much salt. Clean up your samples using a PCR purification kit, DNA clean up kit or ethanol precipitation method to remove excess salts from your samples prior to electrophoresis.
• Your DNA may be contaminated with excess protein.  Remove excess protein from the DNA sample by phenol extraction or DNA purification kit before you run your samples.
• Your DNA loading buffer may be the problem. Try loading DNA with another recipe that was properly made.
• You electrophoresed improperly. Either bad gel running voltage or running buffer was used. Don't allow gel running voltage to exceed more than 20 V/cm. Make sure you keep a temperature of less than 30° C during electrophoresis. Make sure your electrophoresis buffer you use is freshly made and had sufficient buffer capacity. To check the buffer capacity of your running buffer, check the pH in the anode and cathode chambers of the gel apparatus.
• If your DNA is small, during staining steps or incubations in buffer the DNA may have diffused. Make sure you either make your gels containing  ethidium bromide and/or run the gel with ethidium bromide.

{mosloadposition advert3}

Related Mass Spectrometry Articles

• No Related Post