Sequence-dependent quenching of fluorescein fluorescence on single-stranded and double-stranded DNA.

ABSTRACT: Fluorescein is commonly used to label macromolecules, particularly proteins and nucleic acids, but its fluorescence is known to be strongly dependent on its direct chemical environment. In the case of fluorescein-labeled nucleic acids, nucleobase-specific quenching originating in photoinduced charge transfer interactions results in sequence-dependent chemical environments. The resulting sequence specificity of fluorescent intensities can be used as a proximity detection tool, but can also lead to biases when the abundance of labeled nucleic acids is quantified by fluorescence intensity. Here we comprehensively survey how DNA sequences affect fluorescence intensity by preparing permutational libraries containing all possible 5mer contexts of both single-stranded and double-stranded DNA 3' o

SUBMITTER: Lietard J 

PROVIDER: S-EPMC8982050 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature