DNA Staining Method Based on Formazan Precipitation Induced by Blue Light Exposure

Aaron J. Paredes; Hilda M. Alfaro-Vald&eacute;s; Christian A.M. Wilson

doi:10.3791/56528

DNA Staining Method Based on Formazan Precipitation Induced by Blue Light Exposure

JoVE Journal
Biochemistry
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JoVE Journal Biochemistry

DNA Staining Method Based on Formazan Precipitation Induced by Blue Light Exposure

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08:11 min

January 28, 2018

DOI: 10.3791/56528-v

Aaron J. Paredes¹, Hilda M. Alfaro-Valdés¹, Christian A.M. Wilson¹

¹Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas,Universidad de Chile

This method allows selective staining and quantification of DNA in gels by soaking the gel in a SYBR Green I/Nitro Blue Tetrazolium solution and then exposing it to sunlight or a blue light source. This produces a visible precipitate and requires almost no equipment, making it ideal for field use.

The DNA staining protocol has three general steps. The first is to put the gel in a solution of SYBR Green I and nitro blue tetrazolium, also known as NBT. The gel is then stirred for 25 minutes if it is a polyacrylamide gel and an hour if it is an agarose gel.

The last step is to expose the gel to a blue light source such as blue LEDs or sunlight. During this step some precipitate bands appears. These bands have a direct correlation with the DNA concentration and the gel then can be digitalized in an office scanner in order to quantify the DNA.

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