nitrocellulose,nylon and baking

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Mon Nov 11 15:12:05 EST 1991


In article <9111081440.AA14796 at calvin.jci.tju.edu>, buchberg at CALVIN.JCI.TJU.EDU (Dr. Art Buchberg behind the variety of techniques that can be don> Can you uv Xlink Nitrocellulose?
> ... Can you bake nylon, (either charged or uncharged)
> or do you really have to do anyhting for nylon membranes.

> ...Art Buchberg
> 
It looks to me like you would like to know what people have tried and what
works for them.  I baked nylon before UV linking devices came out and it
worked fine.  Now, we have a Stratalinker UV crosslinker and I routinely
crosslink nylon (Duralon UV from Stratagene) and nitrocellulose (Schleicher &
Schuell BA85) with it.  One thing which we do (recommended by Stratagene) is to
place the blot, DNA or RNA side up, on a piece of Whatman paper soaked in 10X
SSC during crosslinking.  Supposedly, this pulls the DNA down onto the blot by
capillary action.  After linking, I soak the blot in distilled water to remove
the salt and go on with hybridization, etc.  Thus, my blots are never dry. 
When I was routinely using a vacuum oven to bake my blots, I used to dry them
first with a heat lamp which was far enough above the blot to prevent a fire!
-Barbara Bugg
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Barbara Bugg, M.S.  bugg at mbcf.stjude.org       "I refuse to have a battle of
St. Jude Children's Research Hospital          wits with an unarmed person."
Memphis,  TN  USA
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