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Different emulsions containing various antioxidants (14 mL) were transferred into the amber bottle and capped tightly.
Liquid samples (0.5 mL) were transferred into sealed vials (15 mL) and incubated for 5 min at 75 °C.
An aliquot (3 ml), were transferred in a 25 ml volumetric flask and diluted with MeOH except of petroleum ether fraction which were diluted to 5 ml.
100 μg mL-1 aliquots of the standard solutions (A: 0.05 0.8 and B: 0.1 1.0 mL) were transferred to a series of 10 mL calibrated flasks.
Plasma samples (1 ml) were transferred to 10-ml centrifuge tubes, and then 20 μl internal standard solution (50 μg/ml) was added.
Approximately 20 ml were transferred to a high-density polyethylene scintillation vial and acidified to pH 2 with trace-metal-grade nitric acid.
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To a set of 10 ml volumetric flasks, aliquots of ebastine standard solutions over the concentration range of 0.1-1.0 0.1-1.0ere transferred, 1 ml of 1.5 × 10-3M eosin were added, followed by 2.5 ml of mcliavain buffer pH 2.2, the volume was completed to mark with distilled water and mixed well.
For the kinetic study, 2.5 ml of EBS standard solution (400 μg/ml) were transferred into a series of 25 ml volumetric flasks to obtain a final concentration of 40 μg/ml, 0.4 M sodium hydroxide or 0.4 M hydrochloric acid were added for method I. Regarding method II, 0.1 M hydrochloric acid or 0.1 M sodium hydroxide were used.
Four mL of heparinized whole blood (10 IU/mL) were transferred to sterile polypropylene conical tubes containing equal volumes (3 mL) of Histopaque-1119 and 1077 (Sigma, St . Louis MO, USA).
The cells at a density of 2.5 × 10 cells/mL were transferred to 1.5 mL centrifuge tube and spun down at 400 × g for 5 minutes at room temperature.
Stably transfected cells grown to 3 × 106 cells/ml were transferred to 500-ml spinner flasks and cultured at 25°C with constant stirring (80 rpm) until a doubling of the cell density with a viability of >90% was reached.
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