A simplified diphenylamine colorimetric method for growth quantification
A simplified diphenylamine colorimetric method for growth quantification
复制标题
用于生长定量的简化二苯胺比色法
DOI：
10.1007/s00253-013-4893-y
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发表时间：
2013-04
影响因子：
5
通讯作者：
Yang, Keqian
中科院分区：
文献类型：
--
作者：
Zhao, Youbao;Xiang, Sihai;Dai, Xida;Yang, Keqian
在生物学研究和生物过程优化中需要监测细胞生长。在特殊情况下，例如在含有不溶性颗粒的培养基中进行微生物发酵，准确的细胞生长定量是当前方法的挑战。在这种情况下，只有伯顿法才适用。原Burton法通过采用高氯酸程序中的两步样品预处理来简化，以消除DNA提取的需要。在这里，我们进一步简化了伯顿法取代以前的两步高氯酸预处理与一个新的和一步二苯胺试剂预处理。通过测定大肠杆菌、棒状链霉菌、酿酒酵母和里氏木霉等四种模式微生物在普通培养基或含有固体颗粒的培养基中的生物量，对该方法的可靠性和准确性进行了评价。结果表明，该方法与传统的OD600法和Burton法相比，具有更高的灵敏度。总的来说，新方法简单、可靠、易于执行，并且在大多数情况下普遍适用，并且它将操作时间从超过12 h（对于先前简化的Burton方法）减少到约2 h。
Cell growth needs to be monitored in biological studies and bioprocess optimization. In special circumstances, such as microbial fermentations in media containing insoluble particles, accurate cell growth quantification is a challenge with current methods. Only the Burton method is applicable in such circumstances. The original Burton method was previously simplified by adopting a two-step sample pretreatment in perchloric acid procedure to eliminate the need for DNA extraction. Here, we further simplified the Burton method by replacing the previous two-step perchloric acid pretreatment with a new and one-step diphenylamine reagent pretreatment. The reliability and accuracy of this simplified method were assessed by measuring the biomass of four model microorganisms:Escherichia coli,Streptomyces clavuligerus,Saccharomyces cerevisiae, andTrichoderma reeseigrown in normal media or those containing solid particles. The results demonstrate that this new simplified method performs comparably to the conventional methods, such as OD600or the previously modified Burton method, and is much more sensitive than the dry weight method. Overall, the new method is simple, reliable, easy to perform, and generally applicable in most circumstances, and it reduces the operation time from more than 12 h (for the previously simplified Burton method) to about 2 h.
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DOI：
10.1016/s0021-9258(17)41524-9
发表时间：
1945-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
G. Schmidt;S. J. Thannhauser
通讯作者：
G. Schmidt;S. J. Thannhauser
影响因子：
3.6
作者：
I. Santamarta;M. T. López-García;R. Pérez-Redondo;B. Koekman;J. Martín;P. Liras
通讯作者：
I. Santamarta;M. T. López-García;R. Pérez-Redondo;B. Koekman;J. Martín;P. Liras
DOI：
--
发表时间：
1946-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
W. C. Schneider
通讯作者：
W. C. Schneider
影响因子：
4.4
作者：
FINDLAY, RH;KING, GM;WATLING, L
通讯作者：
WATLING, L
影响因子：
5
作者：
van Veluw, G. Jerre;Petrus, Marloes L. C.;Claessen, Dennis
通讯作者：
Claessen, Dennis