Assay for Telomerase Activity Using Microfluidic Device
OwnerCalvin B. Harley, Hui Wang +1inventors
InventorsCalvin B. Harley, Hui Wang, Zhu Zhen Pirot
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Abstract
Methods for determining the level of telomerase reverse transcriptase activity in mammalian cells are disclosed. A preferred measuring device is a microfluidic device that includes a spectrophotometer, a fluorescent detector, a fluorescence polarization detector or a scintillation counting device.
Claims20
A method of measuring telomerase repeat amplification products in a cell or tissue reaction mixture comprising flowing the telomerase repeat amplification products in a channel of a microfluidic device and measuring the amount of telomerase repeat amplification products in the reaction mixture.
A method of measuring telomerase activity in a cell or tissue extract, the method comprising the steps of:
- (a) placing an aliquot of cell or tissue extract in a reaction mixture comprising a telomerase substrate and a buffer in which telomerase can catalyze extension of said telomerase substrate by addition of telomeric repeat sequences to generate telomeric extension products which are then amplified to generate telomerase repeat amplification products;
- (b) separating the telomerase repeat amplification products in a channel of a microfluidic device, and
- (c) measuring the amount of telomerase repeat amplification products in the reaction mixture.
A method for evaluating the biological response of a subject exposed to a telomerase modulator comprising:
- (a) obtaining a cell or tissue sample from the subject,
- (b) generating a telomerase repeat amplification product reaction mixture from the subject's cell or tissue sample,
- (c) separating the telomerase repeat amplification products in a channel of a microfluidic device, and
- (d) measuring the amount of telomerase repeat amplification products in the reaction mixture.
A method for evaluating the biological response of a subject exposed to a telomerase modulator comprising the steps of:
- (a) placing an aliquot of cell or tissue extract from the subject after exposure to the telomerase modulator in a reaction mixture comprising a telomerase substrate and a buffer in which telomerase can catalyze extension of said telomerase substrate by addition of telomeric repeat sequences to generate telomeric extension products which are then amplified to generate telomerase repeat amplification products;
- (b) separating the telomerase repeat amplification products in a channel of a microfluidic device,
- (c) measuring the amount of telomerase repeat amplification products in the reaction mixture, and
- (d) correlating the level of telomerase activity in said cell extract with the level of said extended telomerase substrate.
Dependencies are inferred from the claim text.