Simple and Practical Staining of DNA with GelRed in Agarose Gel Electrophoresis
Huang, Q.; Baum, L.; Wei-Ling Fu
Clinical Laboratory 56(3-4): 149-152
2010
ISSN/ISBN: 1433-6510 PMID: 20476647 Document Number: 14927
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.
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