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For all strains, batch cultures were started in defined medium with 40 mM succinate added as sole C- and energy-source, and 0.5 mM ammonium added as N-source.
After adaptation to the new media, batch cultures were started and sampled daily as described below.
Batch cultures were started by inoculation with 10 log growth phase cells into 1 liter glass flasks filled with 750 ml of Modified Bold-3 N medium [ 49] without soil extract.
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Synechocystis cultures were started from plate and incubated for 3 4 days in batch.
Briefly, the cultures were started by collecting adult beetles and parasitised egg batches from several infested eucalypt plantations in the central North Island of New Zealand (38°14′23′′ S 176°13′22′′ E).
Continuous culture Continuous culture was started with a batch culture at styrene concentration in a solution corresponding to the concentration of this compound in a fed later on nutrient medium.
After a batch phase of approximately 24 hours the continuous culture was started at a dilution rate of D = 0.1 h-1 (growth medium flow rate of 150 g h-1).
Batch cultures were carried out as follows.
Biomass and metabolite concentrations in batch and chemostat and batch cultures were determined as described by Guadalupe et al.[ 6].
Continuous cultivation was started after 80 mmol of NaOH had been added to the batch culture (80% of the xylose exhausted or appr. 3.5 g dry weight biomass formed per kg culture).
As it has been previously demonstrated that there is a direct correlation between the initial biomass concentration and the rate of inhibitor conversion [ 11, 29], some of the batch cultures at 39°C were started with lower inoculum size (OD620 ~0.2).
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