DNA quantification.

werner at inw.agrl.ethz.ch werner at inw.agrl.ethz.ch
Thu Apr 30 04:28:44 EST 1998


In article <Pine.SOL.3.96.980427170955.17666B-100000 at post>, Yankit Tam <yktam at aecom.yu.edu> says:
>
>Hi everyone,
>
>I am looking for an accurate DNA quantification method at
>the level of 30 ng/ul DNA.  I have read references on using
>PicoGreen.  Is this the best and most sensitive way to
>quantiate tiny amount of DNA in solution.   Any other
>suggestions?  Thanks in advance for any hints.
>
>Kit
>
>Kit Tam
>Department of Microbiology and Immunology
>Albert Einstein College of Medicine
>Bronx  NY 10461
>USA
>

I do an ethidium-bromide-plate-assay: 

1) Plate preparation: Pour 60 mm petri dishes containing 5ml 0.8% Agarose,
 1x TE, 1mg/ml EtBr. Store at 4° up to one month

2) Prepare standarts using lambda-DNA, make five dilutions to cover the
 range from 5 to 75 ng/ul. Store at -20°

3) Spot 0.5 ul of each standard and of your sample on the plate and allow
 to absorb into the plate for 10 to 15 minutes at room temp. 

4) Compare the standards to your sample using an UV lightbox.

This method is not very exact but for my applications it is enough and I
like it especially because I only need a very little amount of my 
'valuable' samples.


Patricia 

Patricia Werner
Institut für Nutztierwissenschaften
Tannenstrasse 1 / ETH-Zentrum
CH-8092 Zürich
Switzerland




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