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TRAP法是一种常用的测定端粒酶活性的方法,但并不太适合测定木本植物的端粒酶活性.因此以油松针叶为实验材料,对TRAP法进行改进.参照前人的研究通过对比实验证明,在端粒酶提取过程中将PEG8000加入上清液,可获得高效的端粒酶.在此基础上,对PCR先导引选择TS、模板浓度选择100 ng,最终获得油松针叶高质量的端粒酶PCR产物.应用SYBR Green I替代银染液对电泳凝胶进行染色,获得条带清晰、重复性好的电泳结果.通过对油松针叶、银杏叶片、胡杨愈伤组织、沙冬青悬浮细胞等4种木本植物实验材料进行测定,证明该改进的技术体系可能是适合于木本植物端粒酶活性测定的稳定的、灵敏度高的可行方法.图5表1参17
TRAP method is a commonly used method for the determination of telomerase activity, but it is not suitable for determination of telomerase activity in woody plants.Therefore, the method of TRAP is improved by using pine needle as experimental material.Referring to previous studies Through comparative experiments, it was proved that telomerase could be obtained by adding PEG8000 into the supernatant during telomerase extraction.On the basis of this, 100 ng of TS was selected for the PCR primer and the concentration of the template was 100 ng, Leaf high quality telomerase PCR product.Using SYBR Green I instead of silver staining solution to dye the electrophoresis gel to obtain a clear and reproducible electrophoresis results.After the pine needles, ginkgo leaves, Populus californica callus , Ammopiptanthus mongolicus suspension cells and other four kinds of woody plant test material to determine that the improved technology system may be suitable for the determination of telomerase activity of woody plants stable and high sensitivity feasible method Figure 5 Table 1 参 17