Fluorescence based nuclease assay

Details Fluorescence based nuclease assay Fluorescence based nuclease assay

1999-01-20

2000-10-03

Elliott, George C.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

536 231, 536 232, 435 911, 436 94, G01N 3300, C12Q 168, C12P 1934, C07H 2104

Patent

active

061271241

ABSTRACT:
Methods for the determination of nuclease stability of an oligomeric compound and its deletion sequences by capillary gel electrophoresis using laser-induced fluorescence detection (LIF CGE) are provided. Fluorescently labeled oligomeric compounds are treated with one or more agents having nuclease activity resulting in an assay mixture of the original oligomeric compound and its deletion sequences. A diluted aliquot taken directly from the assay mixture is analyzed using LIF CGE. Results of the assay yield quantitative concentrations of the oligomeric compound and each of the deletion sequences. In further embodiments, the invention provides methods for determining the relative binding affinity of one or more oligomeric compounds for a substrate having nuclease activity, and methods for determining the nuclease activity of an enzyme.

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