Detection of DNA with Quant-iT™ PicoGreen® dsDNA Reagent in microplate format
MIB Application Laboratory, Thermo Fisher Scientific, Vantaa, Finland
Introduction
The PicoGreen reagent is an ultrasensitive fluorometric stain used for the quantitation of dsDNA in a solution. In a wide variety of molecular biology applications the detection and quantitation of small amounts of dsDNA is very important. This reagent is much more sensitive than the most commonly used absorbance measurement at 260 nm (A260). When measuring the absorbance, single-stranded nucleic acids and nucleotides interfere with the obtained signal.
The commonly used bisbenzimide dyes of Hoechst, 33258 and 33342, are affected less by interfering substances than the absorbance measurement but the sensitivity is not as good as with PicoGreen. Only one dilution of the reagent is needed for the whole dynamic range instead of two different fluorochrome dilutions which are generally needed with the Hoechst dye.
In combination with Thermo Scientific Fluoroskan Ascent or Fluoroskan Ascent FL, the PicoGreen reagent provides rapid and easy determination of dsDNA, offering a wide dynamic range.