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1a f, Mm a – Eppendorf BioSpectrometer kinetic User Manual

Page 95

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95

Evaluation procedure

Eppendorf BioSpectrometer

®

kinetic

English (EN)

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. 33)
. 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

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