Ethidium bromide stock solution

Ethidium bromide stock solution 10 mg/mL in water, working solution 2.5 pg/mL in water. 

B 1 pg/ml ethidium bromide in TE buffer by dilution of a stock solution of 1 mg/ml EtBr in Soln. A. (The other mentioned dyes are used the same way.)... 

Various size standards are available that should be run together with the samples in order to confirm the expected size of the PCR product. Ethidium bromide (a mutagenic reagent ) is used to visualise the DNA strands it usually sold as a concentrated stock solution (e.g. 10 mg/ml). 

After electrophoresis, transfer the gel into ethidium bromide solution (50 pi stock solution in 1 1 of tap water) and stain for 5 -10 min. Transfer the gel onto a transilluminator and visualise the DNA with UV light, taking care to protect the skin and eyes against UV radiation. Document the result with a standard photograph or digital image analyser. 

Ethidium bromide/acndine orange stock solutions consist of 50 mg of ethidium bromide and 15 mg of acridine orange, dissolved in 1 mL of 95% ethanol and made up to 50 mL with distilled water. Store at -20°C. 

For use in viability counts, stocks are diluted 1.100 with PBS. Stored at room temperature in the dark. The solution has a shelf-life of at least 2 mo. Hazard warning both ethidium bromide and acridine orange have mutagenic properties, are combustible, toxic, and irritants to the skin, eyes, and respiratory system. Handle with care... 

Ethidium bromide solution 10 mg mH stock solution made up of 100 mg of ethidium bromide and 10 ml of water... 

Storage and Handling Ethidium bromide should be handled in the laboratory using the "basic prudent practices" described in Chapter 5.C. Because of its mutagenicity, stock solutions of this compound should be prepared in a fume hood, and protective gloves should be worn at aU times while handling this substance. Operations capable of generating ethidium bromide dust or aerosols of ethidium bromide solutions should be conducted in a fume hood to prevent exposure by inhalation. 

We prepared the stock solutions of acridine orange, proflavin, ethidium bromide, aniline, benzidine and DAPI in a standard buffer containing 8 mmol/L Tils, 50 mmol/L NaCl and 1 mmol/L EDTA, pH 7.0. The stock solutions of naphthalene, anthracene, phenanthrene, parathion, pentachlorophenol, nitrobenzene, naphthalene, benzo[a]pyrene, dibenz[a,h]anthracene, dibenz[a,j]anthracene, benz[a]anthracene, benzoQ]fluoranthene, benzo[k]fluoranthene, and 1,2,3,4,5,6,7,8-octahydronaphthalene were prepared in ethanol. The concentration of the stock solutions for the carcinogens and hydrocarbons was between 1x10" mol/L and 5x10 mol/L. We carried out serial dilutions with the standard buffer before the experimental measurements. Serial dilutions of the stock solutions were prepared in ethanol using standard buffer containing 5% ethanol. 

Highly-polymerized deoxyribonucleic acid, (DNA), from calf-thymus, was purchased from Sigma Chem. Co. and used as received. Stock solutions of DNA were prepared by dissolution overnight in 5 mM phosphate pH 7 buffer and were stored at 4°C in the dark for short periods only. Concentrations of DNA per nucleotide phosphate were determined by absorption spectroscopy using a molar extinction coefficient of 6,600 M cm at 260 nm [4]. N,N -Dimethyl-2,7-diazapyrenium dlchloride (DAP " ") was prepared and purified according to the method of HUnig et al. [5] whereas ethidium bromide (EB ) was purchased from Sigma Chem. Co. and used as received. 

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