Southerns--need help!

Malte Buchholz mbuchho at med.uni-goettingen.de
Tue Mar 30 07:59:07 EST 1999


sokching wrote:
> 
> I'm having problems with my southern blots. The probe is hybridizing to
> everything....therefore a smear appears on the autorad. As I'm using the
> ECL kit, the hybridization temperature cannot exceed 42C. I've tried
> increasing the stringency of the washing but it doesn't help. I'd
> appreciate any ideas. Thanks.
> 
> sokching


Hi sokching,

does your hybridization buffer contain formamide? 50% formamide will
greatly reduce unspecific hybridization at 42°C (I can supply you with a
recipie for the buffer if interested).

Hope this helps,

Malte



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