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Self-Quenched Covalent Fluorescent Dye–Nucleic Acid Conjugates as Polymeric Substrates for Enzymatic Nuclease Assays

✍ Scribed by Vladimir S. Trubetskoy; James E. Hagstrom; Vladimir G. Budker


Publisher
Elsevier Science
Year
2002
Tongue
English
Weight
65 KB
Volume
300
Category
Article
ISSN
0003-2697

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✦ Synopsis


A fluorescent method is described for assessing nuclease activity. The technique is based on the preparation of quenched fluorophore-nucleic acid covalent conjugates and their subsequent dequenching due to degradation by nucleases. The resulting fluorescence increase can be measured by a spectrofluorometer and exhibits subpicogram per milliliter sensitivity level for RNase A and low picogram per milliliter level for DNase I. The method is adaptable for quantitative nuclease inhibitor testing.