Dai Dongsheng, Chen Yi, Qi Li, Yu Xiao
Center for Molecular Science, Institute of Chemistry, People's of Republic of China.
Electrophoresis. 2003 Sep;24(18):3219-23. doi: 10.1002/elps.200305600.
Sharp peaks of Escherichia coli JM 109 (up to 1 300 000 theoretical plates) were recorded with either extremely diluted (< 5 mM) or extremely concentrated (ca. 150 mM) Tris-borate (TB) running buffers. However, under the conditions of yielding sharp peaks, migration time of E. coli was irreproducible. Critial factors influencing reproducibility were found to include bacterial growth phase, storage condition, cell pretreatment before injection, and concentration of running buffer. Buffer concentrations in the range of 20-100 mM TB were essential for reproducibility. E. coli JM 109 was shown to be sensitive to ultrasonification. Bacterial growth and storage conditions could be monitored by CE, with results comparable to those obtained with optical methods.
使用极度稀释(<5 mM)或极度浓缩(约150 mM)的 Tris-硼酸盐(TB)运行缓冲液时,可记录到大肠杆菌JM 109的尖锐峰(理论塔板数高达1300000)。然而,在产生尖锐峰的条件下,大肠杆菌的迁移时间不可重现。发现影响重现性的关键因素包括细菌生长阶段、储存条件、进样前的细胞预处理以及运行缓冲液的浓度。20-100 mM TB范围内的缓冲液浓度对于重现性至关重要。已证明大肠杆菌JM 109对超声处理敏感。细菌的生长和储存条件可用毛细管电泳监测,结果与光学方法所得结果相当。