预冷的1M山梨醇。若用抗性筛选方法，则立即加入1ml冰预冷的YPD/山梨醇。轻柔转移细胞悬液至一个无菌管中。

? 检查和记录脉冲参数。时间常数应接近5ms。电场强度可以通过实际电压（kV）

/电击杯间隙宽度（cm）获得。

? 用营养缺陷突变株进行互补筛选时，细胞应立即涂布至含有1M山梨醇的限定琼脂

平板（minimal agar plate）上。当用抗性筛选方法时，电转后的细胞在30℃振荡培养1-2小时；在含有1M山梨醇的YPD琼脂平板上，用适当的抗生素进行电穿孔细胞的培养。30℃培养72-96小时。

5.3 电转化所需溶液和试剂

? YPD/HEPES：100ml YPD培养基，20ml 1M HEPES，pH8.0。 ? 1M DTT：1.55g DTT，溶于8ml水，定容至10ml，过滤除菌。

? YPD/山梨醇：10g yeast extract，20g peptone，182.2g山梨醇，溶于700ml

水，定容至900ml。高压灭菌。临用前加入100ml无菌20%葡萄糖。

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Troubleshooting Guide for the Micropulser：略 产品信息

系统特点：

? 处理样品的速度更快——简单的一键“Pulse”操作、快速的充电时间。 ? 快速的程序选择——内置的、优化的程序适合细菌和酵母的通用研究。 ? 电弧淬灭系统（ARQ，Arc quenching system）——减少电弧效应，保护有价

值样本的损失。

? 更宽的参数手动优化空间——手动程序允许在200-3000V范围内设置电压，精

度10v；脉冲时间可以在1.0-4.0ms范围内调整，精度0.1ms。 ? 3000V容量——可以提高大体积的转化效率。 ? 紧凑型、节省空间的设计。 ? 可听见和可视化的脉冲指示。

? 可显示时间常数和实际释放电压——可监测重复性。

为什么要用电转化？

电转化是目前可用的效率最高的转化方法。它比化学转化方法的转化效率高几个数量级，

并且比其他方法重复性更好。MicroPulser被设计为E.coli、fungi和其他微生物的电转化提供最佳的电条件，可以获得最高的转化效率。