NanoPhotometer
®
P-Class
User Manual
Version 2.1
Page 68 / 70
Formula for ssDNA:
FOI = (8.77 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320)]
With dye correction:
FOI = (8.77 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320 – CF
dye
* (Abs.
max, dye
– Abs. 320)]
Formula for RNA:
FOI = (8.11 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320)]
With dye correction:
FOI = (8.11 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320 – CF
dye
* (Abs.
max, dye
– Abs. 320)]
Formula for Oligonucleotides:
FOI = (9.83 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320)]
With dye correction:
FOI = (9.83 * (Abs.
max, dye
– Abs. 320) / [ε
dye
* 10
-6
* (Abs. 260 – Abs. 320 – CF
dye
* (Abs.
max, dye
– Abs. 320)]
Abs.
max, dye
absorbance at absorption maximum of the dye (AU)
ε
dye
dye-dependent extinction coefficient (M
-1
* cm
-1
)
The following dye types and parameters are pre-programmed in the NanoPhotometer
®
P-Class.
Dye Type
Absorption maximum Dye
(nm)
Dye-dependent extinction
coefficient
ε
dye
Dye-dependent correction
factor (260 nm) CF
Dye
Alexa Fluor 350
345
18,400
0.25
Alexa Fluor 488
492
62,000
0.30
Alexa Fluor 532
525
82,300
0.24
Alexa Fluor 546
555
104,000
0.21
Alexa Fluor 555
555
150,000
0.04
Alexa Fluor 568
576
93,000
0.45
Alexa Fluor 594
588
80,400
0.43
Alexa Fluor 647
650
239,000
0.00
Alexa Fluor 660
660
107,000
0.00
Alexa Fluor 680
680
164,000
0.00
Cy3
550
150,000
0.08
Cy3.5
581
150,000
0.08
Cy5
649
250,000
0.05
Cy5.5
675
250,000
0.05
Oyster-500
503
78,000
0.29
Oyster-550
553
150,000
0.05
Oyster-556
560
155,000
0.03
Oyster-645
649
220,000
0.05
Oyster-650
653
200,000
0.04
Oyster-656
660
200,000
0.04
Texas Red
593
85,000
0.23
*In all formulas the molar dye-dependent extinction coefficient is used.