Eppendorf BioSpectrometer fluorescence - Manual (Page 93)

93
Evaluation procedure
Eppendorf BioSpectrometer ® fluorescence
English (EN)
For ssDNA, RNA, Oligo:
MM = calculated relative molar mass; unit: kDa
bp = entered number of base pairs per molecule
b = entered number of bases per molecule
12.5.4 Calculating the factor for protein in "General Method Parameter"
This section only covers the calculation of the protein components in the Dye labels and Proteins direct
UV method groups. For these method groups, the protein component is selected in the parameters (see
Method parameters on p. 36). The protein component is assigned a factor that will be entered in the
General Method Parameter/Proteins function for each protein. Alternatively, A
0.1%
or the absorbance
coefficient plus the molar mass of the protein can be entered instead of the factor. In this case, the factor is
calculated as follows:
F = factor for the protein; unit: g/L.
A
0.1%
= absorbance of the protein at a concentration of 0.1 % (1 g/L).
When entering the molar absorbance coefficient and the relative molar mass of the protein A
0.1%
can be
calculated on this basis:
ε
P
= molar absorbance coefficient of the protein; unit: cm -1 M -1 .
MM
P
= relative molar mass of the protein; unit: Da (entry in General Method Parameter in kDa).
• For dsDNA the calculation of the molar concentration is based on the assumption of a
double-stranded nucleic acid. For the ssDNA, RNA and Oligo methods, a single-stranded
nucleic acid is assumed.
• For methods which have been reprogrammed via <New Method> in the Routine main
group, Nucleic acids method group, always double-stranded nucleic acids are assumed
for calculating the molar concentration.
% 1 . 0
1
A
F
P
P
P
MM
A
H
% 1 . 0