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The combined extracts were filtrated and concentrated to remove ethanol.
Alq3-ethanol solvated phase was annealed at ~200°C under nitrogen atmosphere for 2 hrs to remove ethanol from the crystal lattice.
After magnetically stirring for 1 min (constant temperature magnetic mixer, DF-101S, Zhengzhou Greatwall Scientific Industrial and Trade Co., Ltd., Zhengzhou, China), the resultant emulsion was evaporated under vacuum and dark for 30 min at 35 °C to remove ethanol.
To identify root colonization, the roots were washed to remove ethanol, cut into 1 cm length, and insert into the heat resistant bottle containing 10% KOH solution then autoclaved for 20 min at 120 °C.
After filtration through Whatman No. 1 filter paper, litchi pericarp extract (LPE) was concentrated at 50°C using a rotary evaporator under vacuum to remove ethanol, and then adjusted to 500 ml with distilled water.
Cells were fixed with 70% ethanol at RT, 20 30 min, stored at −20°C overnight, centrifuged for 5 min at 200×g to remove ethanol, and washed ×1 in PBS.
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To remove ethanol-traces the columns were centrifuged for an additional minute.
Thus, reaction–separation integration by removing ethanol from the zone where the biotransformation takes place, offers several opportunities for increasing product yield and consequently reducing product costs.
After removed ethanol and added 0.5 ml staining solution (50 µg/ml PI, 100 µg/ml RNaseA, 0.2% Triton-100), cells were incubated in 37°C for 30 min in the dark.
The ethanol extracts were filtrated, concentrated and removed ethanol by reduced pressure.
Tissue was removed, ethanol evaporated to dryness with nitrogen and reconstituted in assay buffer.
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