• Fluorescence is measured at 525 nm following excitation (470 nm) by the BLUE LED.
• Relative fluorescent output at 525 ± 20 nm (user settable cursor).
• Sample concentration is populated based on a standard curve. There are six choices of units.

The PicoGreen® dye is a fluorescent nucleic acid stain for quantitating double-stranded DNA (dsDNA). Used in conjunction with the micro-volume capability of the NanoDrop® ND-3300 Fluorospectrometer, the PicoGreen® assay provides a highly sensitive means of dsDNA quantitation with minimal consumption of sample.

The main disadvantage of general UV spectroscopy for dsDNA quantitation is the contribution of signal from single-stranded DNA (ssDNA) and other contaminants, such as protein and extraction buffers. PicoGreen® reagent circumvents such contributions from interfering substances by exhibiting an emission maximum at 530nm when bound specifically to dsDNA (unbound PicoGreen® reagent exhibits minimal fluorescence in solution). The ability of the ND-3300 fluorospectrometer to measure as little as 1 ul of sample, allows significantly scaled-down reaction volumes, thereby using only a fraction of sample commonly needed for conventional cuvette-based fluorometers.

The ND-3300 fluorospectrometer has demonstrated a detection range for dsDNA bound with PicoGreen® reagent of 1pg/ul - 1000pg/ul, and has proven to be 75 times more sensitive then using the Hoechst 33258 dye with this system. Readings taken at the lowest detection limit consume only 2 picograms of dsDNA per measurement. The following protocol is an adaptation of the Molecular ProbesTM dsDNA Quantitation Reagent and Kits product information sheet.