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After 5 days of cultivation, samples were harvested and biomass formation and cellulase activity were measured as described previously [ 51].
After 24, 48, 72, and 96 h of cultivation, samples were withdrawn, and the activity of 2-methylcitrate synthase was determined.
For preparation of metabolite extracts, the cultivations were sampled two (wild-type experimental design cultivation samples for LC-MS/MS analysis) or three (for all other conditions) times for each of the two MS-methods (biological replicates) to be employed, using fast vacuum filtration and quenching of metabolism in 37.5% cold methanol (-20°C).
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During batch cultivations, culture samples were collected at different time intervals for monitoring growth, and the culture supernatant was collected for analysis of glucose, acetic acid, lactic acid, propionic acid, and ethanol.
From each of the 24 cultivations samples were taken for transcriptome analysis.
During batch cultivations, 5��mL of culture samples were collected in quadruplets when cultures reached stationary phase.
Ethanol of varying concentration was added at the beginning of cultivation. 1 mL of culture samples were took and measured (OD730) every 12 h.
Hexane of varying concentration was added at the beginning of cultivation. 1 mL of culture samples were collected and measured with a spectrophotometer at OD730 every 12 h.
During the 10 day cultivation period, samples of C. vulgaris were collected daily for biomass analysis.
The temperature was maintained at 35.0 ± 0.5 °C throughout the cultivation and samples were collected every 24 h.
The offline analysis of the cultivation broth samples included the determination of the OD600 and the glucose, nitrate and Surfactin concentration of the supernatant.
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