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Background: Although SYBR Gold or SYBR Green I have been used in the loading buffer as a DNA stain safer than ethidium bromide for agarose gel electrophoresis, electrophoretic mobility of DNA is altered and thus DNA fragment size cannot be accurately determined.
Methods: A method using GelRed in the loading buffer was developed to stain DNA fragments in agarose gel elec-trophoresis.
Results: Among various concentrations of GelRed, SYBR Gold, or SYBR Green I tested in the loading buffer, only the highest tested concentration of GelRed, i.e., 100× GelRed, did not change band mobility. Evaluations using various sizes of PCR products at different concentrations further confirmed that 100× GelRed could be used to ac-curately determine DNA fragment size. The reagent can be stored at 4 °C for at least 1 year without a decrease in staining sensitivity.
Conclusions: The 100× GelRed is a sensitive and safe alternative to ethidium bromide and better than either SYBR Gold or SYBR Green I for size determination in agarose gel electrophoresis. Our laboratory now uses the GelRed method routinely with great consistency and success.
DOI: Clin. Lab. 2010;56:149-152
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