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PIPES monosodium salt hydrate, 99%


Company: Alfa Aesar
Catalog#: B21835
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The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" Cells
[Abstract]  This protocol differs from other procedures in that the bacterial culture is grown at 18 °C rather than the conventional 37 °C. Otherwise, the protocol is unremarkable and follows a fairly standard course. Why growing the cells at low temperature should affect the efficiency of transformation is unknown. Perhaps the composition or the physical characteristics of bacterial membranes synthesized at 18 °C are more favorable for uptake of DNA, or perhaps the phases of the growth cycle that favor efficient transformation are extended. Incubating bacterial cultures at 18 °C is a challenge. Most laboratories do not have a shaking incubator that can accurately maintain a temperature of 18 °C summer and winter. One solution is to place an incubator in a 4 °C cold room and use the temperature ... [摘要]  该方案与其它方法的不同之处在于细菌培养物在18℃而不是常规37℃下生长。否则,协议是不明显的,并遵循一个相当标准的课程。为什么在低温下生长细胞应该影响转化的效率是未知的。也许在18℃合成的细菌膜的组成或物理特性更有利于DNA的摄取,或者有利于有效转化的生长周期的阶段延长。在18℃孵育细菌培养物是一个挑战。大多数实验室没有可以准确保持温度在夏天和冬天18°C的摇动孵化器。一种解决方案是将培养箱放置在4℃冷室中,并使用温度控制将培养箱加热至18℃。或者,如果培养物在许多实验室中的环境温度20-23℃下生长,则几乎没有效率损失。在这些温度下温育的培养物以2.5至4小时的倍增时间缓慢生长。为了避免在晚上达到期望的OD,在晚上建立培养物并且在第二天早晨收获细菌。该程序适用于许多菌株。通常用于分子克隆,包括XL1-Blue,DH1,JM103,JM108/9,DH5a和HB101。