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Southern Blot Transfer Buffer Substitutions?


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#1 ickyostes

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Posted 27 May 2009 - 10:46 AM

I have some sodium acetate trihydrate on hand, and am wondering if it would work to substitute a buffer from this for the usual sodium citrate buffer that is used for Southern blot capillary transfers.  The two buffers have really similar pH ranges and are both sodium salts, but I'm wondering if there are other considerations.  I'm using a pretty standard protocol with a vertical capillary transfer from an agarose gel to a nylon membrane.  If any of you have used this, what was your buffer solution?

#2 Carlton H

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Posted 27 May 2009 - 07:07 PM

You should be fine.  The chemistry is very similar.  Just remember that sodium acetate has 1/3rd the ionic strength of citrate, so you'll want to use more.  I want to tell you to just use 3x as concentrated sodium acetate as you would sodium citrate but for some reason I feel like there's something I'm not thinking of.

... I think you should be fine with acetate.

-Carlton
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