Question:
Why is my electrophoresis 14 % gel is wavy?
I need to mark LC3 protein, which has 16-14 kDa. When I do a 14% gel for the electrophoresis after the 20 kDa, my gel bands stay wavy (as shown in the photo) so it is not possible to get the protein.
A wavy gel can be caused by a number of factors, including:
Answer:
Uneven gel polymerization: The gel may not have polymerized evenly, which can cause uneven bands or a wavy appearance. To prevent this, make sure that the gel is allowed to polymerize completely before use, and that the gel is level when it is poured.
Uneven sample loading: Uneven sample loading can cause distortions in the gel. Make sure that you load your samples evenly and accurately.
Insufficient running buffer: Make sure that you are using enough running buffer in the electrophoresis chamber. If the buffer level is too low, it can cause uneven current flow and distortions in the gel.
Overloading of samples: Overloading of samples can cause distortions in the gel, as the samples will not migrate uniformly through the gel. Try reducing the amount of sample loaded onto the gel.
Electrical problems: Make sure that your electrophoresis apparatus is working correctly and that there are no electrical problems that could be causing the gel to wave. Check the power supply, cables, and connections.
Age of the gel: If the gel is old or has been stored improperly, it may not perform as expected. Try using a fresh gel and see if the problem persists.
In addition, the band broadening or waviness in high percentage gels like 14% can also be attributed to the high proportion of cross-linker in the gel which can lead to a more rigid and less flexible gel matrix.
You can try optimizing the electrophoresis conditions, such as the buffer composition, running voltage, and running time. Additionally, you may want to try using a different percentage gel, such as a 12% or 15% gel, to see if this improves the resolution of your protein bands.
Lastly, make sure that you are using a high-quality protein ladder to accurately identify the size of your protein of interest.
#dna #gelelectrophoresis #genetics #pcr #biology
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