ddTRAP: A Method for Sensitive and Precise Quantification of Telomerase Activity
| Autor: | Dawne N. Shelton, Woodring E. Wright, Andrew T. Ludlow, Jerry W. Shay |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Telomerase Polymerase Chain Reaction Article Primer extension 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Line Tumor Neoplasms Biomarkers Tumor Humans Digital polymerase chain reaction DNA Primers Enzyme Assays Chemistry RNA DNA Neoplasm Telomere Reverse transcriptase Cell biology HEK293 Cells 030104 developmental biology Cell culture 030217 neurology & neurosurgery DNA |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493977765 |
| DOI: | 10.1007/978-1-4939-7778-9_29 |
| Popis: | Telomerase is a cellular RNA template-dependent reverse transcriptase that adds telomere repeats to the 3’ ends of chromosomes. Telomerase is expressed almost universally in tumor cells (>85%) to maintain telomere length, thus providing the ability of tumor cells to avoid senescence and to have unlimited replication ability, one of the key hallmarks of cancer. ddTRAP (droplet digital Telomere Repeat Amplification Protocol) is a two-step assay with whole cell lysates that utilizes a telomerase-mediated primer extension followed by droplet digital PCR (ddPCR) detection of extended products. The adoptation of the TRAP assay to ddPCR has resulted in improved throughput, increased sensitivity and better repeatability of the TRAP assay. The protocol described below details our procedures for ddTRAP. |
| Databáze: | OpenAIRE |
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