NanoPhotometer
®
P-Class User Manual
Version 2.1 Page 17 / 70
4.1.4 Dye incorporation for dsDNA, ssDNA, RNA and Oligonucleotides
The dye incorporation methods are similar to the dsDNA, ssDNA, RNA and Oligonucleotide methods. This section
describes the specific features concerning the dye incorporation. For general information please follow the detailed
instructions under Analysis of dsDNA, ssDNA and RNA and Oligonucleotides.
To determine the dye incorporation rate, the absorbance reading at the wavelength reported for maximum absorbance
of the fluorescence dye is used. For further details please refer to 12.2 Nucleic acid fluorescent dye incorporation.
The procedure is as follows:
Parameter Screen
NanoVolume Applications
Cuvette Applications
Parameter Screen
Step 1 Press 1 for NanoVolume OR 2 for Cuvette folder.
Step 2 Press 1 to select Nucleic Acids folder.
Step 3 Press 5, 6, 7 or 8 to select one of the dye incorporation
methods.
Step 4 Using the NanoVolume Applications select the Lid Factor as
described in the “Average Detection Range Sheet” and
under 3.2.
Step 5 Select Dilution Factor, Units and Factor as described under
4.1.2.
Step 6 Select whether the Dye correction (calculation of the dye-
dependent correction factor) is used or not with the left and
right arrows. The Background correction is always
calculated in the Dye methods.
Step 7 Select the appropriate Dye Type. 10 different Alexa Fluors,
4 Cy-Dyes, 6 Oyster-Dyes and Texas Red are programmed
with their corresponding maximum absorbance
wavelength, dye-dependent correction factor at 260 nm
and dye-dependent extinction coefficient. For further details
please refer to 12.2 Nucleic acid fluorescent dye
incorporation.
Step 8 If using Custom Dye maximum absorbance wavelength of
the custom dye, dye-dependent extinction coefficient and
dye-dependent correction factor at 260 nm have to be
entered.
Ranges are:
Dye Abs Max: 300 nm to 950 nm
Dye Ext. Coefficient: 10,000 to 9,999,999
Dye Correction: 0.000 to 0.999
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