The use of 2-aminopurine fluorescence to study DNA polymerase function.

Abstract:

The fluorescence of the base analog 2-aminopurine (2AP) is used in highly sensitive assays to determine kinetic parameters for DNA polymerase catalyzed reactions, including exonucleolytic proofreading and nucleotide binding and incorporation. Since 2AP fluorescence can also be used to probe DNA polymerase-induced conformational changes in 2AP-labeled DNA substrates, reaction steps that occur before product formation can be detected. Instruction is provided here in the use of 2AP fluorescence in steady-state and presteady-state assays to study DNA polymerase function and DNA replication.

Polymerases:

T4

Topics:

Biotech Applications, Nucleotide Analogs / Template Lesions, Nucleotide Incorporation, Methods

Note:

Chapter 21 of Molecular Methods in DNA Replication. Eds. S. Vengrova, JS Dalgaard

Status:

new topics/pols set partial results complete validated

Entry validated by:

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