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A Fluorescent High-Throughput Assay for Double-Stranded DNA: the RediPlate PicoGreen Assay
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2003
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DnaReplicate Fluorescence MeasurementsGeneticsDna AnalysisMolecular BiologyDouble-stranded DnaPicogreen ReagentBioanalysisRediplate Picogreen AssayDna ComputingAnalytical BiotechnologyLaboratory MedicineDna SequencingBiochemistryFluorescent High-throughput AssayMolecular Biological MethodDna ReplicationNatural SciencesFluorescent Picogreen ReagentNucleic Acid AmplificationMedicineGenome EditingHigh-throughput Screening
The fluorescent PicoGreen reagent for detection and quantitation of double-stranded DNA has been adapted for high-throughput screening: the RediPlate PicoGreen double-stranded DNA assay format. In the RediPlate PicoGreen assay format, the PicoGreen reagent is predistributed and co-dried into either 96- or 384-well microplates with the excipient trehalose. The user resuspends the dried reagents upon adding DNA, and measures the resulting fluorescence after a five minute incubation. Replicate fluorescence measurements on nominally identical wells have less than a 5% coefficient of variation. The assay is linear from 5 to 500 ng/ml DNA in a 200 micro l volume. The RediPlate PicoGreen assay format retains the advantages of the original PicoGreen reagent - sensitivity, speed, and specificity - but in a high-throughput format.