Northern Blot - METHYLENE BLUE

Cathy Sprankle Sprankle at ciit.org
Tue Sep 19 00:29:47 EST 1995


In article <43m270$j6n at chaos.kulnet.kuleuven.ac.be>,
dirk.jochmans at chaos.kulnet.kuleuven.ac.be wrote:

(snip)
> Is it possible to stain  a Gene Image Hybound membrane with something to
see the rRNA blotted? I heard something about Methylene blue or Gold
Stain??
> Could this be a good method to verify the amount of RNA (in stead of
using an actine probe).
> 
> Thanks,
>          Dirk
We use Amersham's Hybond N, a neutral nylon membrane, for our Northerns,
and stain them (after UV crosslinking) with a methylene blue solution. 
Combine 45 mls 3M NaAcetate pH 5.3 and 225 mls ultrapure water (doesn't
have to be DEPC water, but I'd want to be resonably sure there had been no
grubby fingers in it).  Add about 5 mg methylene blue, shake/stir till
dissolved.  Soak blot in stain 5 min, destain in water 5 min.  The first
time I tried this, I made two identical blots, stained one but not the
other, and hybridized them both.  The autorads were identical.  (Sorry I
don't have a reference for this, got the method from a colleague.)

Good luck!
Cathy

-- 
Catherine Sprankle
CIIT
e-mail:  sprankle at ciit.org
Opinions expressed (such as they are!) are strictly mine and do
not reflect those of CIIT.



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