Eppendorf BioSpectrometer kinetic - Manual (Page 31)

31
Methods
Eppendorf BioSpectrometer ® kinetic
English (EN)
6.2.2 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.
Nucleic acids
• Determination of the concentration of nucleic acids through measurement at 260 nm and evaluation via
factor.
• Various nucleic acid methods, such as dsDNA or RNA, are preprogrammed. The parameters vary
according to the factor.
• Preprogrammed method for microliter cuvettes: Measuring DNA in sample volumes within the
microliter range with 1 mm light path (with microliter cuvettes as Eppendorf μCuvette G1.0 or Hellma ®
TrayCell).
• Additional information on the purity of the measured nucleic acid: Ratios A260/A280, ratios A260/A230,
absorbance wavelength spectrum of nucleic acid, absorbance of the background wavelength (preset:
320 nm; the absorbance of the pure nucleic acid should be close to zero here).
• Partial turbidity correction can be performed via the Background parameter.
• Concentrations can be converted to molar concentrations and (after the sample volume has been
entered) to nucleic acid quantities ( process results method step).
Proteins direct UV
• Determination of the concentration of proteins via measurement at 280 nm and factor or standard
evaluation.
• Preprogrammed methods for direct absorbance output as a result (Protein A 280) and for evaluation via
albumin-specific absorbance coefficients (Albumin A 280).
• Preprogrammed method for microliter cuvettes: Measuring protein in sample volumes in the microliter
range with 1 mm light path (with microliter cuvettes as Eppendorf μCuvette G1.0 or Hellma ® TrayCell).
• Additional information on the purity of the measured protein: Absorbance of the background
wavelength (preset: 320 nm; the absorbance of the pure protein should be close to zero here).
• Partial turbidity correction can be performed via the Background parameter.
• When programming the methods, the corresponding factor is imported through the simple selection of
the protein from a predefined list. The factors are separately defined in the functions of the Gen.
method param. group. Various proteins are preprogrammed in Gen. method param.; additional
proteins can be added.
Proteins (with reagent)
• Concentration determination of proteins via measurement according to color reactions and evaluation
using standards or factors (typical: evaluation with standard curve).
• The Bradford, Bradford micro, Lowry, Lowry micro, BCA and BCA micro methods are already
preprogrammed. According to the reagent manufacturer, the "Curve fit" (standard curve type) must be
changed as necessary.
Dye labels
• For dye-labeled biomolecules: Concentration determination of the biomolecule (nucleic acid or protein)
via measurement at 260 or 280 nm and measurement of the dye in one measuring procedure.
• Evaluation with factor. In addition to the biomolecule, up to two dyes can be measured at the same time
as two different wavelengths.
• Additional: evaluation of the frequency of incorporation (FOI) of the dye. Selection between two
different FOI calculation procedures.
• Already preprogrammed methods: ssDNA, labeled with Cy 3 or Cy 5.