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Ethidium Bromide Solution Preparation and use

Ethidium Bromide

  • Ethidium bromide is used to visualize DNA in agarose gels and in many cesium chloride gradient protocols. 
  • Ethidium bromide is a toxic chemical and a mutagen. It causes eye and skin irritation.
  • Wear gloves at all times and wash contaminated skin with water. 
  • May be stored indefinitely in a lightproof container at room temperature
  • Ethidium Bromide Appearance: Red to purple powder
  • Molecular formula: C21H20N3Br
  • Molecular weight: 394.3
  • Melting point: 255-266°C with decomposition

Ethidium Bromide Solution use

  • Ethidium bromide is commonly used nucleic acid stain for gel electrophoresis.
  • As a stain to visualize U937 cells to assess cell viability
  • Use to detect polymerase chain reaction products
  • In agarose gel electrophoresis based gel retardation assa.

Components for Ethidium Bromide Solution

The following components are required to prepare 1L of Ethidium Bromide Solution

ComponentAmountConcentration
Ethidium bromide (mw: 394.31 g/mol)10 g0.0254 M

Ethidium Bromide Solution Preparation

  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 10 g of Ethidium bromide to the solution.
  3. Add distilled water until volume is 1 L.
  4. Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved. Wrap the container in aluminum foil or transfer the 10 mg/mL solution to a dark bottle and store at room temperature.

References 

  • “Ethidium Bromide Solution Preparation.” AAT Bioquest, Inc, 26 May. 2021, https://www.aatbio.com/resources/buffer-preparations-and-recipes/ethidium-bromide.
  • https://www.bio-rad.com/webroot/web/pdf/lsr/literature/4006020b.pdf
  • https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Product_Information_Sheet/e1385pis.pdf
  • https://www.sigmaaldrich.com/catalog/product/sigma/e1510?lang=en&region=IN

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