Hello. I have problem in gel electrophoresis on rice DNA after PCR with primers. The bands are blur and fade. Can anyone suggest me the solution? whether it is related to voltage or some reagent or gel preparation.
Hello Ishwinder
There are many points to consider. They are as follows:
1. There could be excess of salts in the sample.
2. There could be a problem in the DNA extraction protocol.
3. The quality of water used in making the buffers.
4. Check on the percentage of agarose gel you are using.
5. Do not run the gel too fast.
6. Change the run buffer frequently.
7. The gel may be heating up during the electrophoresis run. If yes, then try decreasing the voltage.
All the best.
the top bands in lanes 3 and 6 are blurred because the is too much material loaded. The smallest bands in all tracks are blurred because they are small and diffuse easily in the warm gel under high voltage. The middle bands are probably blurred by thermal diffusion because the gel is running too hot. Try running the checker gel less far and at half the voltage that you are using and the blurring should improve
Thanks for your answer Malcolm Nobre. I was using the same material and reagents before with good results and resolution. I started using green taq buffer without mgcl2. However, I added mgcl2 separately @ .9ul per 15ul reactio. I used 180 v to run the samples.
I used distilled water for preparing buffers and autoclaved distilled water for pcr reaction.
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