DNA quantification

Patricia Werner werner at inw.agrl.ethz.ch
Tue Jun 22 03:34:34 EST 1999


DNA below 1 ng/ul seems a very low concentration to me. Maybe a misprint ? If so,
there is a protocol to measure DNA concentrations in the range from  0 to about 100
ng/ul.

1) Pour agarose-ethidium bromide plates (0.8% agarose, 1ug/ml EtBr, 1xTBE). Can be
stored at 4°C up to 1 month
2) Spot 0.5µl lambda-DNA standards  (eg. 100, 75, 50, 25, 10, 5, 0 ng/µl ) and the
samples to be measured onto the plate. Be careful not to dig into the surface of the
plate, instead let capillary action pull the small volume from the pipet tip to the
surface.
3) Allow to dry for 15 minutes at room temperature and compare the samples to the
standards using an UV-lightbox.

Patricia

Kevin Urbanic wrote:

> I would like to know if anybody has a method of determining [DNA] other
> than using a UV-vis spectrophotometer.  Our departmental spec seems to
> provided extremely inconsistent concentrations when I spec plasmid DNA
> that is at a concentration less than 1ug/ml.  Specifcally, I was
> wondering if anybody has calculated the concentration of each band in a
> 1kb DNA ladder?  I thought that could be a useful tool for "eyeing" the
> DNA concentration.
>
> K
>
> --
> ********************************************************************************
>
>                "If you love something set it free
>                 If it comes back to you it is yours
>                 If it doesn't then it was never meant to be"
>
> Kevin Urbanic
> Department of Microbiology
> University of Guelph
> Guelph, ON
> N1G 2W1
> Phone: (519) 824-4120
>
> *******************************************************************************




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