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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