Patent ID: 8182986
Filing Date: 2012-05-22
Classification: C12Q,G01N

Abstract:
1. A method for detecting the effect of a test compound being tested for effect on nuclease activity, said method comprising: incubating in an assay medium said test compound, a nuclease which is either an endonuclease or exonuclease V, and an enzyme donor fragment (ED)-reagent for sufficient time for reaction of said ED-reagent and said nuclease, said ED-reagent comprising ED linked through an oligonucleotide strand of not more than 60 nucleotides to a magnetic bead, said reaction resulting in separating said ED from said magnetic bead; adding enzyme acceptor (EA) and a β-galactosidase substrate that allows for determination of an amount of turnover of said substrate; wherein said ED and EA are fragments of β-galactosidase, and said separating of said ED from said magnetic bead results in complexing of said ED and said EA to form an active β-galactosidase enzyme, and said ED reagent has substantially reduced ability to complex with said EA as compared to ED separated from said bead, thereby resulting in reduced background; and determining the amount of turnover of active β-galactosidase enzyme formed from said ED and said EA as a measure of the effect of said test compound on said nuclease activity.