1. Academic Validation
  2. DNA Electrophoresis Using Thiazole Orange Instead of Ethidium Bromide or Alternative Dyes

DNA Electrophoresis Using Thiazole Orange Instead of Ethidium Bromide or Alternative Dyes

  • J Vis Exp. 2019 Mar 31:(145). doi: 10.3791/59341.
Casey S O'Neil 1 Jacie L Beach 1 Todd D Gruber 2
Affiliations

Affiliations

  • 1 Department of Molecular Biology and Chemistry, Christopher Newport University.
  • 2 Department of Molecular Biology and Chemistry, Christopher Newport University; todd.gruber@cnu.edu.
PMID: 30985761 DOI: 10.3791/59341
Abstract

DNA gel electrophoresis using Agarose is a common tool in Molecular Biology laboratories, allowing separation of DNA fragments by size. After separation, DNA is visualized by staining. This article demonstrates how to use thiazole orange to stain DNA. Thiazole orange compares favorably to common staining methods, in that it is sensitive, inexpensive, excitable with UV or blue light (to prevent sample damage), and safer than ethidium bromide. Labs already equipped to run DNA electrophoresis experiments using ethidium bromide can generally switch dyes with no additional changes to existing protocols, using UV light for detection. Blue-light detection to avoid sample damage can additionally be achieved with a blue-light source and emission filter. Labs already equipped for blue-light detection can simply switch dyes with no additional changes to existing protocols.

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