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Common DNA staining methods include the Feulgen reaction, methyl green–pyronin staining, and acridine orange fluorescence staining. Among these, the Feulgen reaction is the most classical histochemical method for DNA detection.
The principle of the Feulgen reaction is based on mild acid hydrolysis (e.g., hydrochloric acid), which cleaves the glycosidic bonds between purine bases and deoxyribose in DNA and breaks the phosphodiester linkage, generating free aldehyde groups on deoxyribose. These aldehyde groups react in situ with Schiff reagent to form a purplish-red compound, specifically labeling DNA-containing regions. RNA is not hydrolyzed under these conditions; therefore, this method is specific for DNA and not suitable for RNA detection.
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