Thermo Scientific NanoDrop One User Manual page 45

Dye concentrations are calculated from the
absorbance value at the dye's analysis
wavelength, the dye's extinction coefficient,
and the sample pathlength. A sloped-line
dye correction may also be used.
Thermo Scientific
Dye absorbance
• Dye absorbance values are measured at specific
wavelengths. See
wavelengths used.
• If Sloping Dye Correction is selected, a linear baseline is
drawn between 400 nm and 750 nm and, for each dye,
the absorbance value of the sloping baseline is
subtracted from the absorbance value at each dye's
analysis wavelength. Baseline-corrected dye absorbance
values are reported and used to calculate dye
concentrations.
Dye correction
• Pre-defined dyes have known correction values for A260
and A280. See Dye/Chromophore Editor
values used.
• A260 dye corrections are subtracted from the
absorbance value
concentration, and from the A260 absorbance value
used to calculate the
Sample Pathlength
• For micro-volume measurements, the software selects
the optimal pathlength (between 1.0 mm and 0.03 mm)
based on sample absorbance at the analysis wavelength.
• For cuvette measurements, pathlength is determined by
the cuvette Pathlength setting in the software (see
General Settings).
• Displayed spectra and absorbance values are normalized
to a 10 mm pathlength equivalent.
4
Nucleic Acid Applications
Measure Microarray
Dye/Chromophore Editor
used to calculate nucleic acid
A260/A280 purity ratio.
NanoDrop One User Guide
for analysis
for correction
A260
43