Hello,
We’ve run a few SDS page gels in the past few weeks but so far, we have not been getting much bands. We bought a PET-21-b+ plasmid and eventually transformed them. After growing it up in 2xyt and lb, and getting an OD of around 0.7, we followed an SDS page protocol to view Our protein bands. we are using a 12% 30 uL gel and loading about 20 uL of sample into each well.
Some comments based on my bad gel experiences:
1) ensure you are using good beta-2 me to denature your sample
2) ensure that you are using the correct ratio of loading dye to sample
3) do not run the gel top fast
4) some of your wells you are overloading
It looks to me like the gel has not set properly. Try freshly made APS and give the gel longer to set
I would also load far less protein your gel is clearly overloaded. How fast did you run your gel ? Did you cool down the buffer ?
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