help: c-fos probe

Cathy Sprankle Sprankle at ciit.org
Sun Jan 28 21:44:05 EST 1996


In article <4e8sd1$dai at masala.cc.uh.edu>, lhe at dna.bchs.uh.edu (Lihua He) wrote:

> Hi:
> Is there any one have good experience using c-fos probe on Northern blot? I  
> have problem with mine because it has high backgroud binding. 

Have you tried increasing your hybridization temperature?  I got very nice
hybridization to my northerns (poly A+ RNA from rat and mouse liver)
hybridizing at 55 deg. C., usually for 2 to 3 days (although I think
overnight worked also--it's been a while).  Hyb was followed by brief room
temp washes with 2X SSC, 1 wash at 55 deg. C with 1X SSC/0.1% SDS for 30
min and 1 wash at 55 deg. C with 0.1X SSC/0.1% SDS.  I should add that my
probes were labeled with 32-P and I was using nylon membranes.

Good luck--feel free to e-mail me if you have questions.

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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