摘要
为了建立一种便于检测端粒酶活性的方法 ,在Kim等开发的TRAP法的基础上作了一些改进。即把细胞提取液与TS引物混合 ,30℃保温 30min以延伸TS引物之后经过酚 /氯仿抽提、乙醇沉淀 ,再做PCR扩增。PCR产物经 12 %非变性聚丙烯酰胺凝胶电泳 ,用银染法来显示电泳结果。结果表明该方法可以有效去除抑制Taq酶活性的因素 ,得到清晰的 6bp阳性条带 ,具有特异性好、灵敏度高、易操作、无放射性危害等优点。
To analyze telomerase activity by Telomeric Repeat Amplification Protocols,radioautography marked by isotopes is widely used,which is not only harmful to health but also cause pollution in environment.In hope to discover a non-radioactive,easy-to-use and sensitive protocol for the detection of telomerase activity,first the telomerase was extended and the synthesized telomerase extension products were extracted using hydroxybenzene/chloroform,then it was deposited with ethanol,followed by PCR amplification.Silver staining is adoppted to show the results of the electrophoretic analysis of the PCR products.The result of the experiment proves that this protocol make it possible to have perfect 6bp positive stripes.
出处
《生物技术》
CAS
CSCD
2001年第2期4-7,共4页
Biotechnology
