摘要
基于微孔板高通量筛选中氧气供给与混合情况往往是微型化培养的关键,本文利用亚硫酸钠法对微孔板的K_La进行了测定。因K_La易受外界环境的影响,实验考察了摇床振幅、无菌介质、孔板装液量及种类等条件的影响,结果表明:在温度34℃、摇床振幅和转速分别为25 mm和200 r/min、以孔径为0.22μm的疏水性微孔滤膜为无菌介质、装液量为0.9 mL且加有玻璃珠的48孔板的条件下,微孔板的K_La值能达到102.00 h^(-1),在同样条件下摇床上摇瓶的对照结果为103.48 h^(-1)。就供氧能力而言,微孔板完全可以达到常规摇瓶的效果,能够保证高通量培养时高好氧微生物的需氧要求。
Oxygen transfer and mixing were the key factors affecting the high-throughput screening with microplates. The sodium sulfite method was used to determine the KL a of microplate. For the KLa was easily disturbed by the perturbation of cultivation environment, the factors such as the shaking frequency, the decontaminating sterile media, the charge volume of the mieroplate and the formats of the microplate were investigated. The results showed that under the conditions of 34 ℃, 25 mm of the eccentric distance, 200 r/min of the shaking speed, the 0.22 μm of hydrophobic microfilter as the decontaminating media, 0.9 mL of the charge volume and 48-well microplates with beads, the KL a could reach 102.00 h^-1 which was comparable to the Kua of 103.48 h^-1 obtained in the flasks.
出处
《工业微生物》
CAS
CSCD
2010年第2期15-19,共5页
Industrial Microbiology
关键词
KLA
亚硫酸钠法
微孔板
高通量
KLa
sodium sulfite
microplate
high-throughput screening
