Eppendorf BioPhotometer plus - Manual (Page 46)

11 Evaluation procedure
46
BioPhotometer plus — Operating manual
11.5 Special evaluation procedures for nucleic acids and protein UV
This section relates to the evaluation of nucleic acids or proteins in the methods dsDNA, ssDNA,
Oligo DNA, RNA, Oligo RNA, Protein UV and the corresponding biomolecule components in the
dye methods.
11.5.1 Correction A
340
Application: Partial correction of distortions of absorbance due to turbidity in the measuring
solution.
The application of the evaluation procedure can be activated in the parameter list of the method.
A
X, corr
= calculated corrected absorbance at a wavelength of 230, 260 and 280 nm
A
X
= measured absorbance at a wavelength of 230, 260 and 280 nm
A
340
= measured absorbance at a wavelength of 340 nm
11.5.2 Correction A
550/650
Application: Correction of the effect of the dye absorbance on the nucleic acid or protein
absorbance at 260 and 280 nm.
The application of the evaluation procedure can be activated in the parameter list of the method.
A
X, corr
= calculated corrected absorbance at a wavelength of 260 and 280 nm
A
X
= measured absorbance at a wavelength of 260 and 280 nm
KF
X
= correction factor for the wavelength 260 or 280 nm (the two correction factors for 260 and
for 280 nm are programmed in the parameter list of the method)
A
X50
= measured absorbance at a wavelength of 550 (or 650) nm
11.5.3 Conversion into molar concentrations and nucleic acid amounts
The conversion can only be applied to nucleic acids and dye methods with nucleic acids as
biomolecule component.
Calculation of amount
Application: Calculation of the amount (mass) of nucleic acid or nucleic acid dye conjugate in the
whole sample volume.
M = calculated total amount (mass) of the nucleic acid or the conjugate in the sample tube
C = calculated concentration of the nucleic acid or the conjugate
V
P, total
= volume of the sample in the sample tube; entered by the user in the third screen of the
result display (see Nucleic acids on page 19).
Hint!
The absorbance values displayed in the results are the directly measured, not the corrected
absorbance values.
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