The Novel Green Plus provides an easy 2-step method to stain the DNA band from DNA electrophoresis. This unique reagent ensures the DNA to be stained with a high sensitivity and good quality. Novel Green Plus is a next-generation DNA-binding dye ideal for use in quantitative real-time PCR (qPCR) and many other applications. We designed the dye by taking into consideration several essential dye properties relevant to PCR, including PCR inhibition, safety, and stability and fluorescence spectra of the dye. Ethidium Bromide (EtBr), which presents sensitivity for detecting 1-5 ng doublestranded DNA (dsDNA) in the agarose gel analysis, has been the most common dye for nucleic acid gel staining. However, several drawbacks of EtBr have been understood, including that EtBr is a mutagen / carcinogen and presents a high risk of inducing cancer. Furthermore, the ultraviolet (UV) light used to illuminate EtBr-DNA compounds probably results in skin or eye damage to the user if misconduct. It’s also noted that exposure to the UV light might cause chemical modifications of the DNA samples in the gel, such as the formation of TT dimmers, leading to challenge with the subsequent DNA manipulations. The Novel Green Plus shows a high sensitivity bound with DNA (Fig.1 ), it also brings a more reliable and safer experience of use, since the stained gel can be visualized with the blue-light transilluminator, thus avoiding the risk of skin/eye damage as well as reducing the side effects of DNA modification caused by the UV light.
Fig. 1 The 1 KB DNA Ladder (250-10,000 base pairs, DM010-R500, GeneDireX) was 2X serial diluted (from 1 to 128 dilution, and the concentration of the red mark is 0.72 ng/ 5ul) and loaded in the 1% agarose gel.
After electrophoresis, the gel was stained with Novel Green Plus for 10 minutes. The gel was observed the blue-light transilluminator.
Novel Green Plus is excited at 497 nm but also shows a secondary excitation peak at 248 nm (Fig.2a). After bound to DNA, the fluorescent emission of the Novel Green Plus is centered at 524 nm (Fig. 2b). These spectral characteristics enable this fluorescent dye to be compatible with a wide variety of gel reading facilities.