What does a TRAP assay measure?
Abstract. The telomeric repeat amplification protocol (TRAP) assay is a highly sensitive PCR based assay and is an important tool for understanding the role of telomerase in cancer. This assay measures an enzymatic activity where the amount of target is dependent upon the activity of the enzyme.
What is TRAP PCR?
The TRAP assay is a popular method to determine telomerase activity in mammalian cells and tissue samples (Kim et al., 1994). The TRAP assay includes three steps: extension, amplification, and detection of telomerase products. NT is its own reverse primer, which is not a substrate for telomerase.
Does PCR use telomerase?
The telomere repeat amplification protocol (TRAP) for the human reverse transcriptase, telomerase, is a PCR-based assay developed two decades ago and is still used for routine determination of telomerase activity.
How is telomerase activity measured?
Telomerase activity can be measured in vitro by using the telomeric repeat amplification protocol (TRAP) (4). This assay has been used extensively to study telomerase activity in uncultured and cultured samples of normal and tumor tissue from many cell types.
What is trap staining?
The TRAP Staining Kit (cat. #AK04) is used for the staining of tartrate-resistant acid phosphatase in osteoclasts. Bone mass is controlled by the balance between the activity of osteoblasts and the activity of osteoclasts. The TRAP Staining Kit supplements the Bone Reabsorption Assay Kits.
How do you measure telomeres?
These over-the-counter telomere length tests use a tool called quantitative polymerase chain reaction (qPCR). The method pools the cells in a patient’s blood sample, extracts their DNA, amplifies the DNA and measures the amount of repetitive DNA sequences found within the DNA pool.
What are telomeres made of?
A telomere is the end of a chromosome. Telomeres are made of repetitive sequences of non-coding DNA that protect the chromosome from damage. Each time a cell divides, the telomeres become shorter. Eventually, the telomeres become so short that the cell can no longer divide.
What is the difference between telomeres and telomerase?
Telomeres function by preventing chromosomes from losing base pair sequences at their ends. Telomerase, also called telomere terminal transferase, is an enzyme made of protein and RNA subunits that elongates chromosomes by adding TTAGGG sequences to the end of existing chromosomes.
How does a telomere function?
Their job is to stop the ends of chromosomes from fraying or sticking to each other, much like the plastic tips on the ends of shoelaces. Telomeres also play an important role in making sure our DNA gets copied properly when cells divide. This shortening is thought to be one of several factors that causes cells to age.
What cell types is telomerase active in?
Telomerase is active in gametes and most cancer cells, but is normally absent from, or at very low levels in, most somatic cells.
How is trap used to determine telomerase activity?
The TRAP assay is a popular method to determine telomerase activity in mammalian cells and tissue samples (Kim et al., 1994). The TRAP assay includes three steps: extension, amplification, and detection of telomerase products.
What are the steps in the TRAP assay?
The TRAP assay includes three steps: extension, amplification, and detection of telomerase products. In the extension step, telomeric repeats are added to the telomerase substrate (which is actually a non telomeric oligonucleotide, TS) by telomerase.
Which is the reverse primer used to amplify telomerase?
TSNT is, an internal standard control, amplified by TS primer. NT is its own reverse primer, which is not a substrate for telomerase. These primers are used to identify false-negative results by if the gel lacks internal control bands. Materials and Reagents
How does telomere play a role in aging?
This results in continuous proliferation of cells which is a hallmark of cancer. Therefore, telomere biology has a central role in aging, cancer progression/metastasis as well as targeted cancer therapies.
