How to get dna quantities from an agarose gel ?

[Eddie Iannuccelli]

Hello,
I am trying to find the easiest way to get dna quantities from agarose
gels bands, the gel provide one or more quantities control bands. The
ImageJ gel menu method return only areas  that are not corrected by
calibration, isn' it ?. Currently I first calibrate image using 3 or 4
control bands (which function best fit dna quantification ?) then I
manually select bands before measuring them  (or threshold  + particle
analyzer if bands intensity are not too heterogeneous) in order to get
the Integrated Intensity which represent the dna quantity.
Do you know a better method or perhaps  a plugin that do that better?

Regards

******************************************
Eddie Iannuccelli
Laboratoire de génétique cellulaire
INRA - Castanet Tolosan
Tel: [phone] / Fax: [phone]

[Phase GmbH]

Hi,

you should make shure that there are no saturated pixels in your bands. In
case of Ethdium Bromide fluorescence (bright bands) avoid pixel values 255
(in case of 8 Bit imaging device, B/W). You may use a LUT to colorize
saturated pixels.

Moreover, your imaging device should work linearly, that means display of
gray values in linear dependence to light intensity. In case of CCD cameras,
the camera gamma should be 1.0.

Hope this helps

Christian Kreutzfeldt