Abstract
Acridine orange is a nucleic acid binding dye that emits green fluorescence when bound to double-stranded DNA or RNA and red fluorescence when bound to single-stranded DNA or RNA under ultraviolet light. This unique characterization allows it to be used for distinguishing or visualization of dsRNA. Here, we present a convenient and efficient protocol for detecting dsRNA in polyacrylamide gels.
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References
Palmgren MG (1991) Acridine orange as a probe for measuring pH gradients across membranes: mechanism and limitations. Anal Biochem 192(2):316–321
Hitoshi Y, Lorens J, Kitada SI, Fisher J, LaBarge M, Ring HZ, Francke U, Reed JC, Kinoshita S, Nolan GP (1998) Toso, a cell surface, specific regulator of Fas-induced apoptosis in T cells. Immunity 8(4):461–471
Wainwright M (2001) Acridine-a neglected antibacterial chromophore. J Antimicrob Chemother 47(1):1–13
Browning CH, Cohen JB, Gaunt R, Gulbransen R (1997) Relationships between antiseptic action and chemical constitution with special reference to compounds of the pyridine, quinoline, acridine and phenazine series. Proc R Soc Lond Ser B 93(653):329–366
Von Bertalanffy L, Bickis I (1956) Identification of cytoplasmic basophilia (ribonucleic acid) by fluorescence microscopy. J Histochem Cytochem 4(5):481–493
McMaster GK, Carmichael GG (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci 74(11):4835–4838
Lillie RD, Conn HJ, Stotz EH, Emmel VM (1977) H. J. Conn’s biological stains: a handbook on the nature and uses of the dyes employed in the biological laboratory, 9th edn. Williams and Wilkins, Baltimore
Plemel JR, Caprariello AV, Keough MB, Henry TJ, Tsutsui S, Chu TH, Schenk GJ, Klaver R, Yong VW, Stys PK (2017) Unique spectral signatures of the nucleic acid dye acridine orange can distinguish cell death by apoptosis and necroptosis. J Cell Biol 216(4):1163–1181
Jacobs BL, Langland JO (1996) When two strands are better than one: the mediators and modulators of the cellular responses to double-stranded RNA. Virology 219(2):339–349
Ablasser A, Bauernfeind F, Hartmann G, Latz E, Fitzgerald KA, Hornung V (2009) RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III–transcribed RNA intermediate. Nat Immunol 10(10):1065–1072
Carmichael GG, McMaster GK (1980) The analysis of nucleic acids in gels using glyoxal and acridine orange. Methods Enzymol 65(1):380–391
Bruno JG, Sincock SA, Stopa PJ (1996) Highly selective acridine and ethidium staining of bacterial DNA and RNA. Biotech Histochem 71(3):130–136
Pichlmair A, Schulz O, Tan CP, Rehwinkel J, Kato H, Takeuchi O, Akira S, Way M, Schiavo G, Reis e Sousa C (2009) Activation of MDA5 requires higher-order RNA structures generated during virus infection. J Virol 83(20):10761–10769
Lauretti F, Lucas de Melo F, Benati FJ, de Mello Volotão E, Santos N, Linhares RE, Nozawa C (2003) Use of acridine orange staining for the detection of rotavirus RNA in polyacrylamide gels. J Virol Methods 114(1):29–35
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Ma, T., Zhao, Y., Cheng, X. (2024). Detection of dsRNA by Acridine Orange Staining. In: Cheng, X., Wu, G. (eds) Double-Stranded RNA. Methods in Molecular Biology, vol 2771. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3702-9_2
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DOI: https://doi.org/10.1007/978-1-0716-3702-9_2
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