NanoPhotometer
®
NP80/N60/N50/C40
User Guide Version 3.1
50
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
��
With background correction without dye correction:
FOI =
324.5 *
(A
max, dye
−
A
BKG
)
ε
dye
* 10
6
* (A
260
−
A
BKG
) *
ε
nuc
Without background correction and with dye correction:
FOI =
324.5 *
A
max, dye
ε
dye
* 10
6
*
(A
260
−
cf
dye
* A
max,dye
) *
ε
nuc
Without background correction and without dye correction:
FOI =
324.5 * A
max, dye
ε
dye
* 10
6
* A
260
*
ε
nuc
FOI
Frequency of Incorporation (dye per 1,000 bases)
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
dye
Extinction coefficient of dye in M
-1
*cm-
1
ε
nuc
Extinction coefficient/nucleic acid factor in ng*cm/µl
cf
dye
Dye-dependent correction factor at 260 nm
