Thermo NanoDrop One, User Manual, Page: 54 / 312

Measure Oligo DNA or Oligo RNA
48 NanoDrop One User Guide Thermo Scientific
Related Topics
• Instrument Settings
Detection Limits for Oligo DNA and Oligo RNA Measurements
The lower detection limits and reproducibility specifications for the oligonucleotide sample
types (ssDNA and RNA) are provided
here . The upper detection limits are dependent on the
upper absorbance limit of the instrument and the extinction coefficients for the user-defined
base sequences.
To calculate upper detection limits for nucleic acid samples
To calculate upper detection limits in ng/μL, use the following equation:
(upper absorbance limit
instrumen
t
* extinction coefficient
sample
)
For example, for a sample measurement using an extinction coefficient of 55, the equation
looks like this:
(550 AU * 55 ng-cm/μL) = 30,250 ng/μL
• Molecular Weight of oligo calculated from user-defined base sequence.
• Number of Bases entered.
• Molar Ext. Coefficient (260 nm) . Molar extinction coefficient of oligo (in
ng-cm/μL) at 260 nm calculated from entered base sequence.
• %GC . Percentage of guanine and cytosine residues in total number of bases
entered.
Baseline Correction On or off
Enter baseline
correction wavelength
in nm or use default
value (340 nm)
Corrects for any offset caused by light scattering particulates by subtracting
measured absorbance at specified baseline correction wavelength from absorbance
values at all wavelengths in sample spectrum. As a result, absorbance of sample
spectrum is zero at specified baseline correction wavelength.
Tip : If the sample has a modification that absorbs light at 340 nm, select a
different correction wavelength or turn off Baseline Correction.
Setting Available Options Description
Note For measurements with 10 mm pathlength cuvettes, the upper absorbance limit is
1.5 AU, which is approximately 75 ng/μL for dsDNA.