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.
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.
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