Simple and practical staining of DNA with GelRed in agarose gel electrophoresis.
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 electrophoresis.


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., 100x GelRed, did not change band mobility. Evaluations using various sizes of PCR products at different concentrations further confirmed that 100x GelRed could be used to accurately determine DNA fragment size. The reagent can be stored at 4 degrees C for at least 1 year without a decrease in staining sensitivity.


CONCLUSIONS
The 100x 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.
Huang
Qing
Q
Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chongqing, China.
Baum
Larry
L
Fu
Wei-Ling
WL
D052160:Benzothiazoles; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D003959:Diamines; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004996:Ethidium; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C474470:SYBR Gold nucleic acid gel stain; C098022:SYBR Green I; D004247:DNA; D004996:Ethidium