Methylene Blue staining

Dennis J. Templeton djt2 at po.CWRU.Edu
Wed Aug 4 16:37:28 EST 1993


In a previous article, fzestabr at bullwinkle.ucdavis.edu () says:

>Brian Halligan (halligan at post.its.mcw.edu) wrote:
>
>: I have heard that Northern Blots can be stained with methylene blue before
>: hybridization to obviate the need for EtBr staining.  Does anyone have a
>: protocol or opinions on this?
> 
>Dear Brian
>
>I found this method but have not tried it yet. I ran across it but neglected
>to write the reference.
>
>Staining of rRNA bound to membranes
>Immerse membrane briefly in 0.03% methylene blue, 0.3 M NaOAc pH 5.2
>Destain in water
>Hybridize as usual
>
We have used this method happily in the past and it works well.

However, Nitrocellulose cannot be used because it binds the dye too 
strongly.  We have used nylon and that is ok and I don't know about the
other membranes.  When we tried the NC, the blot looked like a piece of
blue denim, but the hybridization looked ok.

good luck, 

dennis



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