Herbert Brittney-Shea, Shay Jerry W, Wright Woodring E
The University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Curr Protoc Cell Biol. 2003 Nov;Chapter 18:Unit 18.6. doi: 10.1002/0471143030.cb1806s20.
This unit describes techniques to analyze telomeric length and telomerase activity in human cells. Telomere length can be determined by a modification of Southern blotting in which the analysis of chromosome terminal restriction fragments (TRFs) provides the average lengths of all telomeres in a cell population. Telomerase activity can be measured in vitro by a sensitive and efficient polymerase chain reaction (PCR)-based detection method, also known as telomeric repeat amplification protocol (TRAP). These assays can be used to study the in vitro cellular effects of aging and cancer treatments on telomere biology and telomerase activity.
本单元介绍了分析人类细胞中端粒长度和端粒酶活性的技术。端粒长度可以通过对Southern印迹法进行改进来确定,其中对染色体末端限制片段(TRF)的分析可提供细胞群体中所有端粒的平均长度。端粒酶活性可以通过一种灵敏且高效的基于聚合酶链反应(PCR)的检测方法在体外进行测量,该方法也称为端粒重复序列扩增协议(TRAP)。这些测定可用于研究衰老和癌症治疗对端粒生物学和端粒酶活性的体外细胞效应。