Northern hybridisation

Cathy Aversa aversa at bms.com
Mon Dec 11 13:48:46 EST 1995


In article <DJ6AD4.BvJ at liverpool.ac.uk>, davyjaw at liverpool.ac.uk (Mr D.J.A.
Wilson) wrote:

> Can anyone help me? I am looking for advice on which method is best for
> hybridising a DNA random-prime labelled probe in a northern blot. I cannot 
> seem to get the blots to work although the RNA seems OK and the probe is 
> labelling OK so I think it is going wrong at the hybridisation or washing step.
> Any advice would be a great help. THanks.
> PS. What is the best membrane for northern blots?


I use Nytran (neutral-charge) membrane from S&S with the following
hyb.buffer:
50% formamide/5x SSPE/2% casein (Hammerstein grade) /1% SDS/100-500 ug/ml
ss carrier DNA at 42oC.  I do many Northerns that are hyb'd with various
random primed and end labeled probes, and these hyb. conditions work
consistently well for me.
Hope this helps!

Cathy Aversa



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