Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Detection of DNA in Agarose Gels by Staining

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DNAs that have been separated by migration through agarose gels may be detected by staining with dyes with low intrinsic fluorescence, a strong affinity for DNA, and a high quantum yield of fluorescence after binding to nucleic acids. The greater the increase in quantum yield, the higher the ratio of signal to noise. Bands of DNA stained with these dyes are visualized by illuminating the gel with UV light at one wavelength and recording at another. Methods are described here for staining and visualization of DNA in gels using three dyes: ethidium bromide, SYBR Gold, and SYBR Green 1. Further information on these and other dyes may be found in Chapter 1, in the Introduction to Quantifying DNA.


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