How to Avoid a Leaky SDS-PAGE gel

Because it's happened to all of us -- despite following the protocol, the gel leaks. What did you do wrong? Usually there's just a seal problem, and Dan shows some tricks that you can use to avoid a leaky gel.

Got your own trick? Leave a comment!

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• labtricks
• science
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• techniques
• tips
• tricks
• SDS
• PAGE
• sodium
• dodecyl
• sulfate
• polyacrylamide
• gel
• electrophoresis
• leaky
• parafilm
• biochemistry
• molecular
• biology
• proteins
• video
• tutorial

[alabonneheure1]

Yesterday I tried the parafilm trick and the pen trick at the same time and the darn gel still leaked. I don't know what else to do. Could you post the recipe for the agarose gel trick? I would appreciate it. Thanks so much.

[labtricks]

@alabonneheure1 Make sure you are using fresh ammonium persulphate solution - if it's old, the gel will not polymerize (or will polymerize much slowly), increasing the chances of leaking.

Also, @parugoelbiologist just replied about the agarose trick: add a 4-5% agarose solution to the foam, then put the glass plates on top and let it solidify. Once solid, continue with resolving as usual. Good luck!

[sakaewan]

Try petroleum jelly to seal the bottom of the plate. Works everytime, espectially with apparatus which more difficult to set than BD ^_^

[labtricks]

@sakaewan Thanks for the trick! Right, the pen trick is limited to this apparatus, but petroleum jelly should work for all casting sets. Thanks =)

[labtricks]

@parugoelbiologist Thanks for sharing this! I've never used the agarose trick but will give it a try =)

[parugoelbiologist]

@labtricks agarose/agar gel (abt 4-5%) may be used for the purpose. add the agarose gel to the foam underneath the plates, not like you do with the resolving gel. in molten state, this gel will automatically rise in between the plates. then let it solidify.

[sakaewan]

Try petroleum jelly to seal the bottom of the plate. Works everytime, espectially with apparatus which more difficult to set than BD ^_^

[veetube1414]

Yeah thanks for this! I will try it. I just spent 1 hour repeating it! Urr! Thanks

[labtricks]

@beeryya interesting! thanks for sharing =) could you provide more details about you use that? do you add the agarose gel to the foam underneath the plates, or do you put it in between the glass plates before the resolving gel?

[beeryya]

you can use agarose gel to stop it from leaking