DNA staining in agarose and polyacrylamide gels by methyl green |
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Authors: | P. Murgai M.U.S. Sachdeva R. Das N. Varma |
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Affiliation: | Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India |
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Abstract: | ABSTRACTMethyl green (MG) is an inexpensive, nonproprietary, traditional histological stain for cell nuclei. When bound to DNA and upon excitation with orange-red light, it fluoresces brightly in the far red region. We compared MG with ethidium bromide (EtBr), the conventional stain for DNA in gels, and Serva DNA stain G? (SDsG), a proprietary stain marketed as a safer alternative to EtBr for staining of electrophoresed DNA bands in agarose and polyacrylamide gels. DNA-MG fluorescence was recorded and 2.4 μg/ml MG produced crisp images of electrophoresed DNA after incubation for 10 min. Stain solutions were stable and detection limits for faint bands as well as relative densitometric quantitation were equivalent to EtBr. MG, EtBr and SDsG cost 0.0192, 0.024 and 157.5 US cents/test, respectively. MG is an effective stain for visualizing DNA in agarose and polyacrylamide gels. Its major advantages including low cost, comparable quality of staining, storage at room temperature, photo-resistance and low mutagenic profile outweigh its disadvantages such as staining of tracking dye and requirement for a gel documentation system with a red filter. |
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Keywords: | DNA binding ethidium bromide far red fluorescence gel documentation system intercalating agent methyl green molecular biology Serva DNA stain G? |
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