Quantifying minute amounts of DNA?

Virginia Dress dress at biosci.arizona.edu
Thu Mar 3 18:52:19 EST 1994


In article <2kd3t1$rp3 at eldborg.rhi.hi.is>, hbriem at rhi.hi.is (Helgi Briem
Magnusson) wrote:
> 
> Hello Netters.
>    Does anyone have a protocol for measuring the amount of
> DNA when you only have a tiny amount available, i.e. too
> little to visualize on an EtBr-agarose gel and too little
> to give a reading at 280nm when diluted?
>    Any help would be much appreciated.
> 
> Regards, Helgi Briem
> Inst. Exp. Pathol.
> Keldur, Iceland

I tried quantifying DNA years ago with a fluorimeter and the Hoechst dye
and
like someone else said it was very unreliable.  My problem was the baseline
drift of the fluorimeter, it was an old fluorimeter.  Sounds like things
haven't
gotten any better.  A260/280 has actually worked very well for me,
0.5-1ul/ml
has given good readings (I wish we had smaller cuvettes or were able to
adjust
the reading window size on the spec).
	Good luck.
																G. Dress
																	Biochemistry, U of AZ, Tucson



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