Agarose gel - DEPC-induced fragility? - RNA gels (Jan/31/2006 )

Hi all

I am running RNA on 1.2% agarose gels (WITHOUT formaldehyde, since I glyoxylate the RNA).

I added the agarose to water and then added DEPC, shook, autoclaved, then made up to volume with 10x BPTE buffer. I could see that the gel was unusually fragile when I removed the comb. By the time I did the Northern transfer, it was in several pieces. Has anyone else had this experience? Last time, I did not DEPC-treat the agarose, and the gel was fine.

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I'm not sure my post will help, but...

when my FA gel was fragile, I just changed the agarose into other brand, and the gel became fine.

Why don't use DEPC treated DW before making gel?

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I use DEPC treated DW to make my gel, and I use 1.5% instead of 1.2% agarose gel. I don't know if the % of the gel matters much but definitely use ready-made and autoclaved DEPC distilled water. It makes things simpler.

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