NanoPhotometer
®
N120/NP80/N60/N50/C40 User Manual Version 4.3.1
67
C Concentration in ng/µl
A
260
Absorbance at 260 nm (10 mm path)
A
BKG
Absorbance at selected background wavelength (10 mm path)
A
max,dye
Absorbance value at the absorbance maximum of the dye (10 mm path)
ε
nuc
Extinction coefficient/nucleic acid factor in ng*cm/µl
cf
dye
Dye-dependent correction factor at 260 nm
Đ Manual dilution factor
Note: Dye correction is only available for single dye selection.
Dye Concentration
For dye-labeled nucleic acids, the concentration of the dye is calculated using a modified
form of the Beer-Lambert equation. For these calculations, the instrument considers the
absorption maximum of the dye, the dye-specific extinction coefficient (see Table 2 on page
68). The dye concentration is calculated with or without background correction as follows:
With background correction:
C =
A
max,dye
* Ð
ε
dye
* 10
6
Without background correction:
C =
A
max,dye
*
Đ
ε
dye
* 10
6
C Concentration in ng/µl
A
max,dye
Absorbance value at the absorbance maximum of the dye (10 mm path)
A
BKG
Absorbance at 320 nm (10 mm path)
ε
dye
Extinction coefficient of dye in M
-1
*cm-
1
Đ Manual dilution factor
Frequency of Incorporation (FOI)
FOI is the degree of labeling based on dye incorporation in a labeled nucleic acid sample. It
is generally expressed as the number of dye molecules incorporated per 1000 nucleotides.
FOI can be calculated with or without background/dye correction as follows:
With background correction with dye correction:
FOI =
324.5 * (A
max, dye
A
BKG
)
ε
dye
* 10
6
* A
260
A
BKG
cf
dye
*
A
max,dye
A
BKG
* ε
nuc
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