Eppendorf BioSpectrometer fluorescence - Manual (Page 35)

35
Methods
Eppendorf BioSpectrometer ® fluorescence
English (EN)
6.3 Fluorimetry method description
6.3.1 Routine method group
The methods for the Routine group are preprogrammed as fixed methods. Therefore, a new method name
is required after the method parameters in the fixed preprogrammed methods have been modified.
The following preprogramed methods are based on Invitrogen's standard operating procedure for the
corresponding reagent. The execution of this procedure may require a license from Molecular Probes, Inc.,
Eugene, OR, USA or Invitrogen Corporation, Carlsbad, CA, USA.
Nucleic acids
Fluorimetric concentration determination of nucleic acid after reaction with reagents.
• Measuring DNA with PicoGreen, evaluation with standard curve/line.
• Measuring RNA with RiboGreen, evaluation with standard curve/line.
• Measuring oligonucleotides with OliGreen, evaluation with standard curve/line.
Versions of the method program have been programmed as "short methods". In "short methods",
measurements can only be completed with two standards (zero standard and one other standard). The
results are not as accurate as measurements with several standards, but the accuracy is sufficient for many
purposes because the standard curve (relationship between the measuring signal and the concentration) is
approximately linear.
• Measuring DNA with Qubit reagents, evaluation with standard curve/line.
No "short method" because the standard curve is nonlinear.
Proteins
Fluorimetric concentration determination of proteins after reaction with reagents.
• Measuring proteins with NanoOrange, evaluation with standard curve/line.
The methods is based on Invitrogen's standard operating procedure for this reagent.
No "short method" because the standard curve is nonlinear.
Additional information on sample preparation and execution is available from Eppendorf. The Application
Support contact information can be found on the rear page of this operating manual.
6.3.2 Basic method group
Raw fluorescence
• Measurement of the RFU value.
• A method for measuring at the emission wavelength of 520 nm is preprogrammed.
Standard
• Measurement of the RFU value and standard evaluation.
• A method for standard evaluation is preprogrammed.
The methods with Qubit reagents deviate from Invitrogen's standard operating procedure.
Two additional standard dilutions need to be created.