We report herein a luminescent switch-on label-free G-quadruplex-based assay for the rapid and sensitive detection of polymerase proofreading activity using a novel iridium(III) complex as a G-quadruplex-selective probe. The interaction of the iridium(III) complex with the G-quadruplex motif facilitates the highly sensitive switch-on detection of polymerase proofreading activity. Using T4 DNA polymerase (T4 pol) as a model enzyme, the assay achieved high sensitivity and selectivity for T4 pol over other tested enzymes.
Keywords: ACN; Exo I; Exo III; G-quadruplex; Iridium(III) complex; Luminescent; PCR; Polymerase; Proofreading; T4 DNA polymerase; T4 pol; T7 Exo; T7 exonuclease; Taq DNA polymerase; Taq pol; acetonitrile; calf-thymus DNA; ctDNA; dAMP; dGMP; dNTP; deoxyadenosine monophosphate; deoxyguanosine monophosphate; deoxyribonucleoside triphosphate; double-stranded DNA; dsDNA; exonuclease I; exonuclease III; polymerase chain reaction; single-stranded DNA; ssDNA.
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