Northern Blotting despair!

Dom Spinella dspinella at chugaibio.com
Thu Aug 14 13:11:17 EST 1997


> Does anyone have a reliable Northern Blotting method that will work 
> using total or messenger RNA and probing for low abundance sequences??
> 
> As you can tell we are having rather a terrible time with northern 
> blotting.
> 
> thanks

Is the problem with the RNA electrophoresis, the blotting, or the
probing?  A little more info will be of use.  Also, I wouldn't even
attempt a Northern Blot for a low abundance sequence using total RNA. 
mRNA is only ~3% of the total RNA and it is difficult to use sufficient
amounts of total RNA to permit detection of the low abundance sequence
component (which can represent as little as 0.003% of the total RNA by
weight) without overloading the gel.  Good luck.  -- D.G. Spinella



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