Bio-Rad Experion DNA Analysis Kits - Manual (Page 35)

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The algorithm used to calculate DNA concentration of a sample peak involves the following steps:
1. The DNA concentration per peak area is first estimated using the concentration and peak area of the
internal upper marker.
[DNA]
est
of sample peak = Sample peak area • [upper marker]/marker area
2. Because the ratio of DNA concentration to peak area depends on peak size, a peak injection
correction factor is applied. The [ladder peak]/ladder peak area ratios for the DNA ladder peaks that
encompass the sample peak are used to calculate the correction factor.
Conc. (ng/µl) = [DNA]
est
of sample peak • correction factor
5.3 Performing Concentration Determination
Experion software also automatically performs quantitation of all peaks in a sample by a single-point
calibration to the upper marker. Results are presented in the following columns of the Results tab:
n
Conc . (ng/µl )
n
Molarity (nmol/l)
To view these default relative quantitation results, double-click on a peak in the electropherogram or
right-click on a band in the virtual gel to highlight the data for that peak in the Results tab.
The amount of dye incorporated into the DNA fragment generally increases linearly with size.
The actual composition of the DNA fragment can affect the amount of dye that is incorporated;
this is particularly evident with smaller DNA fragments.
3. The concentration is then converted to molarity using the following equation (660 is an average
atomic mass per bp):
Molarity (mol/L) = Conc.(ng/µl) / (660 • peak size in Kbp) • 10 6
Experion DNA 1K and DNA 12K Analysis Kits