Could you explain to me how TRAP assay works? I want to detect telomerase activity of...

Could you explain to me how TRAP assay works? I want to detect telomerase activity of breast cancer cell after the cell treated with anti-tumor (curcumin).

Expert Solution

The principle of TRAP assay
The TRAP assay is based on telomerase activity. Telomeres are present at the linear end of chromosomes and generally surrounded by proteins. TRAP assay is used to measure the telomerase activity in mammalian cells and tissue samples. It basically consists of three steps extension, amplification, and detection of telomerase activity.
Extension step: In this step telomeric repeats are added to the telomerase substrate.
Amplification: In the amplification step the telomerase product is amplified by Polymerase chain reaction (PCR) using specific upstream and downstream primers.
Detection phase: The presence or absence of telomerase is confirmed by analyzing gel electrophoresis. To avoid false positive results an internal standard control is used TSNT which is amplified by TS primers.
This principle can be extrapolated to study the telomerase activity of breast cancer after treating the cell with curcumin.

gladiator answered 2 years ago

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