用于评估端粒酶活性和端粒长度的非放射性检测方法。
Non-radioactive assay methods for the assessment of telomerase activity and telomere length.
作者信息
Banerjee Partha P, Jagadeesh Shankar
机构信息
Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, Washington, DC, USA.
出版信息
Methods Mol Biol. 2009;523:383-94. doi: 10.1007/978-1-59745-190-1_25.
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. This protocol consists of two steps: first, telomeric repeats are added to the substrate by the enzyme and second, the extended products will be amplified by Taq-DNA polymerase. The amplified TRAP assay products will be separated on 10% native PAGE and detected by SYBR Green I dye.
摘要
端粒重复序列扩增法（TRAP）检测是一种基于聚合酶链反应（PCR）的高灵敏度检测方法，是了解端粒酶在癌症中作用的重要工具。该检测方法测量一种酶活性，其中靶标的量取决于该酶的活性。该方法包括两个步骤：首先，通过该酶将端粒重复序列添加到底物上；其次，延伸产物将通过Taq-DNA聚合酶进行扩增。扩增后的TRAP检测产物将在10%非变性聚丙烯酰胺凝胶上进行分离，并用SYBR Green I染料进行检测。
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