NanoPhotometer
®
N120/
NP80/N60/N50/C40
User Manual Version 4.3.1
68
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
Table 2. Dye Types, Absorbance Max, Extinction coefficient, and dye-dependent
correction factors
NanoPhotometer
®
Model
Dye Type
Absorbance
maximum of
Dye (nm)
Dye-dependent
ext. coeff.
ε
dye
in M
-1
*cm
-1
Dye-dependent
correction factor
at 260 nm cf
dye
NP80/N60/N50/C40
Alexa Fluor 350
345
18,400
0.25
NP80/N60/N50/C40
Alexa Fluor 488
492
62,000
0.30
NP80/N60/N50/C40
Alexa Fluor 532
525
82,300
0.24
NP80/N60/N50/C40
Alexa Fluor 546
555
104,000
0.21
NP80/N60/N50/C40
Alexa Fluor 555
555
150,000
0.04
NP80/N60/N50/C40
Alexa Fluor 568
576
93,000
0.45
NP80/N60/N50/C40
Alexa Fluor 594
588
80,400
0.43
NP80/N60/C40
Alexa Fluor 647
650
239,000
0.00
NP80/N60/C40
Alexa Fluor 660
660
107,000
0.00
NP80/N60/C40
Alexa Fluor 680
680
164,000
0.00
NP80/N60/N50/C40
Cy3
550
150,000
0.08
NP80/N60/N50/C40
Cy3.5
581
150,000
0.08
NP80/N60/C40
Cy5
649
250,000
0.05
NP80/N60/C40
Cy5.5
675
250,000
0.05