Interactions of Janus Green B with double stranded DNA and the determination of DNA based on the measurement of enhanced resonance light scattering.

Abstract

A novel assay of DNA with a sensitivity at the nanogram level is proposed based on the measurement of enhanced resonance light scattering (RLS) signals resulting from the interaction of Janus Green B (JGB) with DNA. At pH 6.37 and ionic strength < 0.20, the RLS signals of JGB were greatly enhanced by DNA in the region of 300-650 nm characterized by three peaks at 416.0, 452.0 and 469.2 nm. The binding properties were examined using a Scatchard plot based on the measurement of the enhanced RLS data at 416.0 nm at a high JGB: DNA molar ratio (R > 2.22), and an aggregation mechanism of JGB in the presence of DNA at the nanogram level is proposed. Linear relationships can be established between the enhanced RLS intensity and DNA concentration in the range 0-3.5 micrograms ml-1 for both calf thymus DNA (ctDNA) and fish sperm DNA (fsDNA) if 2.0 x 10(-5) M JGB is employed. The limits of determination were 8.7 ng ml-1 for ctDNA and 9.9 ng ml-1 for fsDNA, respectively. Synthetic samples were analysed satisfactorily.