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US 5,629,154 AGranted
Telomerase activity assays
AssigneeGeron Corporation
InventorsNam W. Kim, Calvin B. Harley, Scott L. Weinrich
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Abstract
Telomerase activity in a sample can be measured using a two reaction protocol involving telomerase substrate and primer extension steps.
Claims23
A method for determining whether a cell sample contains telomerase activity, said method comprising the steps of:
- (a) preparing a cell extract from said cell sample;
- (b) placing an aliquot of said cell extract in a reaction mixture comprising a telomerase substrate lacking a telomeric repeat sequence and a buffer in which telomerase can catalyze extension of said telomerase substrate by addition of telomeric repeat sequences;
- (c) adding to said reaction mixture a primer comprising a sequence sufficiently complementary to a telomeric repeat to hybridize specifically thereto under conditions such that if an extended telomerase substrate is present in said reaction mixture, said primer will hybridize to said extended telomerase substrate and extend to form a complementary copy of said extended telomerase substrate, thereby forming duplex DNA molecules comprising an extended telomerase substrate bound to an extended primer;
- and
- (d) correlating presence of telomerase activity in said cell sample with presence of duplex DNA molecules comprising an extended telomerase substrate bound to an extended primer and absence of telomerase activity in said cell sample with absence of said duplex DNA molecules.
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Dependencies are inferred from the claim text.