NanoPhotometer
®
N120/NP80/N60/N50/C40 User Manual Version 4.3.1
66
nucleic acid samples can be calculated with or without background correction depending on
enabled/disabled background correction parameter.
Without background correction:
C = A
260
* ε
nuc
* Đ
With background correction:
C =
A
260
- A
BKG
* ε
nuc
* Đ
C Concentration in ng/µl
A
260
Absorbance at 260 nm (10 mm path)
A
BKG
Absorbance at selected background wavelength (10 mm path)
Đ Manual dilution factor
ε
nuc
Extinction coefficient/nucleic acid factor in ng*cm/µl
Table 1. Nucleic acids extinction coefficients (ε
nuc
)
calculated via extinction coefficient of constituent nucleotides entered
calculated via extinction coefficient of constituent nucleotides entered
Option to enter any factor between 15 and 150 ng*cm/µl
Dye-labeled Nucleic Acid Concentration
For dye-labeled nucleic acids, the concentration of the nucleic acid is calculated using a
modified form of the Beer-Lambert equation. For these calculations, the instrument
considers the absorption maximum of the dye and a certain dye-specific correction factor at
260 nm (see Table 2 on page 68). The concentration of a dye-labeled nucleic acid is
calculated with or without background/dye correction as follows:
With background and with dye correction:
C =
A
260
A
BKG
cf
dye
*
A
max,dye
A
BKG
* ε
nuc
* Đ
With background and without dye correction:
C =
A
260
A
BKG
* ε
nuc
* Đ
Without background and with dye correction:
C =
A
260
cf
dye
* A
max,dye
* ε
nuc
* Đ
Without background and without dye correction:
C = A
260
* ε
nuc
* Đ
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