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SYBR Gold staining of single stranded nucleic acids is strongly influenced by the presence of guanosines

Rien, J.; Zimmermann, M.; Hall, J.; Zavolan, M.; Mittal, N.

2026-05-26 molecular biology
10.64898/2026.05.19.722025 bioRxiv
Show abstract

SYBR Gold is one of the most strongly fluorescent members of the SYBR family of asymmetrical cyanine dyes commonly used for staining double and single-stranded DNA and RNA. The dye fluorescence increases dramatically upon nucleic acid binding. The primary mode of binding to double-stranded DNA is through intercalation between the base-paired deoxyribonucleotides, while the precise mode of binding to single-stranded DNA and to RNA is less well understood. Here we show that the fluorescence activity of SYBR Gold is strongly influenced by the presence of guanines in single-stranded nucleic acids. We expect this insight to aid in the evaluation of SYBR Gold-based measurements of nucleic acid abundance and spark further studies into the binding behaviour of the SYBR family dyes.

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