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layer in between resolving and stacking gel

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#1 ahmadfathi

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Posted 28 January 2016 - 01:02 AM

Hi,

 

i just wondering, sometimes i prepare gel, there is a layer forming in between resolving and stacking gel. i used isopropanol as linearize the resolving gel.

Is this the effect of isopropanol? is this can affected the protein migration to resolving gel?

 

Thank you,

AFM



#2 mdfenko

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Posted 28 January 2016 - 07:50 AM

after the resolving gel is fully polymerized, do you flush out the isopropanol well and carefully remove water droplets prior to stacking?

 

if the resolving gel is not fully polymerized (or close enough) then it may continue to "release" a water (or buffer) layer between the resolving and stacking gels.

 

tip: we use isobutanol to overlay the resolving gel because it will not mix in with the resolving gel solution. isopropanol can mix with the gel solution.


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