Counterion-dye staining method for DNA in agarose gels using crystal violet and methyl orange

Abstract

Sensitive and safe methods for visualization of DNA in agarose gels are described. 0.001% crystal violet dissolved in distilled water was used for DNA staining on agarose gels and it could detect as little as 16 ng of DNA (3 kb, pGem-7Zf/EcoRI) without destaining procedure. The detection limit is four times lower than that of ethidium bromide. To improve the sensitivity, we studied a counterion-dye staining method using methyl orange as a counterion-dye which contributes to reduce excessive background staining by crystal violet. Dye concentration, pH of staining solution, mixing molar ratio of two dyes, and staining times were optimized for the counterion-dye staining. By the staining with a mixed solution of 0.0025% crystal violet and 0.0005% methyl orange in distilled water, 8 ng of the 3 kb DNA in an agarose gel was detected within 30 min.

White Light-Activated Antimicrobial Paint using Crystal Violet

“…Crystal violet (CV), toluidine blue O (TBO), and methylene blue (MB) dyes, which are widely used in histological stains and for bacterial classification 41,42 , are also known LAAAs showing photobactericidal activity in white light. The dyes generate singlet oxygen ( 1 O2) and ROS when they are exposed to the light source 43 , and the 1 O2 and ROS cause adverse effects in bacteria such as loss of membrane integrity, inactivation of enzymes and DNA damage resulting in cell death [44][45][46] .…”

Vitamin C-silver: an Environmentally Benign Choice for DNA Visualization on Polyacrylamide Gels

“…The ratio of acrylamide to Bis was 29:1. Two-fold serial dilutions of pUC18 DNA/MspI Makers per DNA fragment bench were loaded onto gel lanes: 1000, 500, 250, 125, 64, 32, 16,8,4, and 2 pg, respectively. Electrophoresis was carried out using Miniprotean III dual slab cells (BioRad, Hercules, CA) and a power PAC 300 (BioRad) at 20 mA/gel, with the TBE buffer used as the running buffer.…”

“…Visible dye-based stain, e.g. methylene blue, 2 brilliant cresyl blue, 3 crystal violet, 4 and nile blue, 5,6 is a low cost and simple method, but is insensitive and time-consuming. Fluorescent stain is the most popular method in use, promoted by recent technological developments in fluorescence methodologies and instruments, such as ethidium bromide, SYBR green, and SYBR gold.…”

A New Organic Dye-Based Staining for The Detection of Plant DNA in Agarose Gels

“…[4,5] Staining DNA with a visible dye is a plausible alternative to conventional fluorescent staining. Several protocols for detecting DNA in agarose or polyacrylamide gels using visible dyes, such as methylene blue, [6,7] brilliant cresyl blue, [8] crystal violet, [9,10] and ethyl violet, [11] have been described previously. [12,13] These methods are simple and safe but require long staining times and provide low sensitivities.…”