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目的针对脉冲场凝胶电泳(PFGE)操作规范(Pulse Net China)为指导的质控考核,优化实验条件,从而得到高质量的实验结果。方法在充分了解实验原理的基础上,以脉冲场凝胶电泳操作规范(Pulse Net China)为指导,对部分实验条件进行优化和效果评价,总结出一套稳定、科学、准确的实验方法。结果通过反复实验,对一系列的实验条件优化后,得到了高质量的实验结果并顺利通过质控考核。结论黄金琼脂糖(SKG)凝胶浓度,酶切时间、电泳时间、胶块脱色时间的选择对结果图谱影响较大,是沙门菌PFGE实验的关键环节。
Objective To optimize the experimental conditions based on the pulse quality assessment guided by Pulse Net China, and to obtain high quality experimental results. Methods Based on the full understanding of experimental principle and under the guidance of Pulse Net China, some experimental conditions were optimized and evaluated, and a set of stable, scientific and accurate experimental methods were summarized. Results After repeated experiments, a series of experimental conditions were optimized to obtain high-quality experimental results and successfully passed the quality control examination. Conclusion The gel concentration of gold agarose (SKG), enzyme digestion time, electrophoresis time, plastic block decolorization time of the choice of a greater impact on the results of the map is the key to Salmonella PFGE experiments.