SDS PAGE experiment had a weird result. Are there any ideas as to why the gel turned out like this?
Could it be that the gel was very dense or the acrylamide mixture was poured unevenly resulting in the bands shown? Or is it something else altogether?
The one that I am most confused is the wavy pattern at the bottom. Does anyone know what could have caused it? Thank you.
Presence of detergents (except SDS) or high concentrations of salts in your sample can result in band distortion. Also wrong pH or too high voltage setting resulting in increased current and heat development may be reasons.
For trouble shooting see here:
https://de.slideshare.net/nishodhsaxena/trouble-shooting-of-sds-page-analysis.
Peter
Hi Ong, if the lane on the left is MW standards, then there is something seriously wrong with the gel preparation. The wavy pattern at the bottom is the least of your problems.
In short: This is a horrible gel and I would put the blame on the reagents used to make the gel.
If you can, buy premade gels, it will be cheaper in the long run.
If you cant do that, buy premade stock acrylamide and stock buffer solutions for making gels.
Hi all, thank you for your suggestions so far.
@Michael Clarke-Whitte I do not think there was any contamination but I could have been careless without knowing, so I'll take that into consideration.
@ Peter H. Roos Hmm, it could have been high concentrations of salts in the sample which will explain the distortions. But I am not too sure about the effects of pH, and if the voltage was too high (It was at 150V). But none of those seem to explain the wavy pattern below. Finally, based on the troubleshooting website you provided, the most similar looking one (in my opinion) would be slide 6 which is uneven pouring of the acrylamide mixture. However, I could not find any slide explaining the pattern below. I have also checked this website http://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html but was unable to find any similar results.
@ Steingrimur Stefansson Yes, the lane on the left is the MW standards and I know that something went wrong with the gel preparation. What I want to know is if there is a specific/possible explanation for the distortions and the wavy pattern at the bottom.
Once again, thank you for your suggestions. I appreciate the help, and look forward to your reply if you got more insights about this result.
Do you make your own gel reagents from scratch? If so, then I would suggest that you buy readymade reagents.
The wavy pattern on the bottom of the gel can be just a byproduct of a bad gel.
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