摘要: 凝胶迁移(EMSA)实验在理论上较简单,但在实际操作中需要对试验测定系统进行优化,缓冲液的选择是需要优化的因素之一.通过用T7RNA聚合酶与T7启动子DNA的结合反应来选择合适的缓冲液,结果表明,不同的缓冲液会影响凝胶迁移试验的结果,5×Buffer1结合缓冲液的效果较好;TBE电泳缓冲液比TAE、TGE的电泳效果好,同时在种不同浓度的TBE缓冲液中,0.2×的TBE效果最好.
关键词:
凝胶迁移,
缓冲液,
条件优化
Abstract: The test of Electrophoretic Mobility Shift Assay is very simple in theory.But to successfully carry out gel shift experiments,it needs to optimize the parameters.Through a combination of T7 RNA polymerase and T7 Promoter for testing proper buffer was selected.The experimental results showed 5 × binding-Buffer 1 was more suitable for the test;TBE running2buffer was better than TAE and TGE buffer;0.× TBE buffer was the best concentration amang there electrophoresis buffer.
Key words:
EMSA,
Buffer,
Optimize
中图分类号:
郑海霞, 王玉国, 冯智富. 不同缓冲液对凝胶迁移试验的影响[J]. 华北农学报, 2009, 24(6): 178-180. doi: 10.7668/hbnxb.2009.06.036.
ZHENG Hai-xia, WANG Yu-guo, FENG Zhi-fu. Effect of the Different Buffer on Electrophoretic Mobility Shift Assay (EMSA)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(6): 178-180. doi: 10.7668/hbnxb.2009.06.036.