Eppendorf AF2200 Plate Reader - Manual (Page 59)

Predefined methods
Eppendorf ® PlateReader AF2200
English (EN)
59
6.2.4 Nucleic acid quantification (Fluorescence 485/535 nm with standards)
Introduction
PicoGreen is a highly sensitive fluorescent nucleic acid stain for the quantitation of double-stranded
(ds)DNA in solution. The measured fluorescence intensity signal is directly proportional to the amount of
dsDNA present in the sample.
RiboGreen is one of the most sensitive stains for the detection and quantitation of RNA in solution. The
measured fluorescence intensity signal is directly proportional to the amount of RNA present in the sample.
By using two or more nucleic acid standards of known concentrations, the nucleic acid concentrations of
unknown samples can be obtained.
Default measurement parameters
• Excitation wavelength: 485 (20) nm
• Emission wavelength: 535 (25) nm
• Number of flashes: 25
• Settle time: 0 ms
• Mode: Top
• Lag time: 0 μs
• Integration: 20 μs
• Gain: ‘optimal’
• Plate definition: Eppendorf Microplate 96/U-PP, black or comparable plates
User defined method parameters
• The measurement is performed at 485/535 nm with default measurement parameters.
• Define the number of blank replicates (possible number: 1 – 8).
• Define the number of standards (possible number: 2 – 12).
• Define number of standard replicates (possible number: 1 – 8).
• Define the concentrations of each standard (Press triangle button to open the menu).
• Define the number of samples (max. 93, depends on number of blanks and standards).
• Define the unit of your concentration.
• Check if entered parameters are correct.
• Required plate layout: Pipette in a vertical order, starting with blanks at position A1. Standards are
pipetted in subsequent wells, samples must be placed in the following wells.
• Press Start .
• For method results and data evaluation principle see chapter method results and evaluation procedure .
The method is restricted to 96-well microplates.