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SDS-PAGE Gel Electrophoresis Forum SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


Air bubbles in PAGE

Air bubbles in PAGE - SDS-PAGE Gel Electrophoresis Forum

Air bubbles in PAGE - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


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  #1  
Old 07-06-2010, 11:55 PM
Pipette Filler
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Default Air bubbles in PAGE



Hi all,
I have problems with air bubbles between gel and the glass plate while casting PAGE. I am not able to figure out the reason for this . Do these air bubbles interfere with protein movement in the PAGE and if it does, can anyone suggest me a way to avoid air bubbles?

Thanks

Silpi
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Old 01-04-2011, 06:15 AM
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Default Re: Air bubbles in PAGE

Hi Silpi,

Try adding a 200 uL layer of Absolute 100% Alcohol after loading your gel. This will eliminate the Oxygen from the gel whilst it's polymerizing. After your gel has set, decant the alcohol and rinse with running gel overlay.

tshego
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Old 06-10-2011, 08:22 PM
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Default Re: Air bubbles in PAGE

I also found saturated butanol works very well ( use as directed above!!).
here is a recipe I use:
Combine 50 ml of n-butanol or isobutanol (or isopropanol) and 50 ml of H2O in a glass bottle and shake. Use the top phase to overlay gels. Store at room temperature indefinitely.
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Old 07-20-2011, 01:00 PM
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Default Re: Air bubbles in PAGE

air bubbles can affect the movement of protein since when the protein get into contact with the bubble, they revolve around the bubble before continuing to move.position the silage slightly on top of the plate in a slanting position to avoid air from getting inside
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