DNA quantification

Chris LaRosa plarosa1 at navix.net
Tue Jun 22 23:18:54 EST 1999


There are several alternatives to quantitating DNA.

Ethidium bromide fluorescence,   Picogreen fluorescence, Hoescht Dye
fluorescence, all requireing a fluorimeter.. I uses these in my lab
routinely.     They are generally more sensitive than UV absorbance.

On a budget you can spot the dna with any of these dyes with the correct
filters and take a photograph and quantitate by eye.   Since everyone has a
uv box and filters you can spot ethidium bromide stained DNA on saran wrap
for instance.

you can also quantitate by using lambda markers for instance in agarose gel.
One calculates the amount    as follows: guess the quantity of the band of
interest relative to the nearest marker band.     This amount x (size maker
band)/total lambda size) x amount load in maker lane.

There are also little dna quantiating sticks sold by various venders,
although I have never used them.   All these methods are discussed in the
molecular cloning manuals.

Dalia wrote:

> http://www.fermentas.lt/fermentas/Profiles/MarkersPr.htm
> You can use MasRuller DNA ladder
> J.Gagilas
>
> Kevin Urbanic <kurbanic at uoguelph.ca> wrote in message
> news:7kb26p$2bv$1 at testinfo.uoguelph.ca...
> > 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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