Visualization of DNA in agarose gels as migrating colored bands: Applications to laboratory techniques
- Univ. of Michigan, Ann Arbor, MI (United States)
We have developed a method to visualize DNA without the use of ethidium bromide and UV radiation. Anionic dyes (colored anion) have long been used in the detection of pharmaceutical amines via ion pairing, here we show that cationic dyes may be used to detect DNA. In gel electrophoresis in which DNA is traveling toward the positive electrode and a cationic dye is traveling toward the negative electrode, we expect ion pairing of the DNA and the dye as they meet in the gel. The dye should bind to the anionic DNA. If the DNA is not completely neutralized by the dye, it should continue to migrate. Ethidium bromide, which is believed to stain DNA primarily by intercalation between bases, exhibits the fluorescence through its cation and also may bind to DNA, to some extent, through ionic pairing. We observed that DNA forms colored bands during electrophoresis in standard agarose gels when a cationic dye is present in the gel and running buffer. DNA in amounts equal to or greater than 80 ng is seen as a discrete migrating colored band in ambient room lighting. Colored bands may be transferred to nitrocellulose by vacuum transfer in room temperature gel dryer, Xeroxed, fixed with NaOH and dye removed with dilute detergent. Also, isolation of DNA bands from preparative gels may be accomplished without the typical use of ethidium bromide and UV radiation which are known to alter DNA and pose hazards to laboratory personnel. We are presently investigating the general utility of using dyes to visualize DNA for various laboratory techniques.
- OSTI ID:
- 134376
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-1110
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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