Herbert Brittney-Shea, Hochreiter Amelia E, Wright Woodring E, Shay Jerry W
Department of Medical and Molecular Genetics, Indiana University Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana 46202-5251, USA.
Nat Protoc. 2006;1(3):1583-90. doi: 10.1038/nprot.2006.239.
The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay has been modified for real-time, quantitative PCR analysis. Here, we describe cost-effective procedures for detection of telomerase activity using a fluorescent-based assay as well as by using real-time PCR. These modified TRAP assays can be accomplished within 4 h (from lysis of samples to analysis of telomerase products).
端粒重复序列扩增 protocol（TRAP）是用于分析细胞或组织提取物中端粒酶活性的两步法。该灵敏检测方法的近期改进包括使用荧光标记引物而非放射性标记引物来消除放射性。此外，TRAP检测已针对实时定量PCR分析进行了改进。在此，我们描述了使用基于荧光的检测方法以及实时PCR检测端粒酶活性的经济高效程序。这些改进的TRAP检测可在4小时内完成（从样品裂解到端粒酶产物分析）。
