Abstract
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 μl volume. The RediPlate PicoGreen assay format retains the advantages of the original PicoGreen reagent - sensitivity, speed, and specificity - but in a high-throughput format.
Keywords: high-throughput, fluorescence, dna, microplate, dye-binding assay, contaminant tolerance
Combinatorial Chemistry & High Throughput Screening
Title: A Fluorescent High-Throughput Assay for Double-Stranded DNA: the RediPlate PicoGreen Assay
Volume: 6 Issue: 4
Author(s): Robert Batchelor, David Hagen, Iain Johnson and Joseph Beechem
Affiliation:
Keywords: high-throughput, fluorescence, dna, microplate, dye-binding assay, contaminant tolerance
Abstract: 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 μl volume. The RediPlate PicoGreen assay format retains the advantages of the original PicoGreen reagent - sensitivity, speed, and specificity - but in a high-throughput format.
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Cite this article as:
Batchelor Robert, Hagen David, Johnson Iain and Beechem Joseph, A Fluorescent High-Throughput Assay for Double-Stranded DNA: the RediPlate PicoGreen Assay, Combinatorial Chemistry & High Throughput Screening 2003; 6 (4) . https://dx.doi.org/10.2174/138620703106298536
DOI https://dx.doi.org/10.2174/138620703106298536 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
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