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The heptane layer was transferred to clear glass tubes and stored at −20°C for 24 h.
At 7 days post germination, seedlings were transferred to clear, sterile boxes containing 50 mL of Hoagland's liquid medium.
Test sample was then transferred to clear and transparent 96-well plates and was read using a microplate reader (Sunrise, Tecan, Switzerland) at 517 nm.
Following incubation for 15 min at 20°C in the dark, samples were transferred to clear polystyrene vials (Sarstedt, Nümbrecht, Germany), 50 μl 0.01 mM coelenterazine was added, and bioluminescence was assayed at 20°C using a MiniLumat LB9506 luminometer (Berthold, Bad Wildbad, Germany).
A fraction (150 μL) of each culture was transferred to clear polystyrene 96-well flat bottom plates (Corning), cells were pelleted by centrifuging plates at 3000 g for 5 min, supernatant containing excess BV was removed, and whole cell fluorescence (λex = 684 nm; λem = 695 720 nm) was measured using a Tecan M1000 plate reader.
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After 1 day, brains were transferred to clearing solution.
A549 and L132 cells were incubated in RPMI containing 10 μM JC-1 at 37°C for 15 min, washed with PBS, and transferred to a clear 96-well plate.
These cultures were transferred to a clear 96 well flat bottom polystyrene tissue culture plates (BD-Falcon, USA) using 200 µl culture/well.
After the third transfer, 100-µL portions of ten-fold serial dilutions of the growth was transferred to optically clear 96-well polystyrene plates (Corning) with each well containing 100-µL cereal grass medium without the supplemental wastewater (CGM) and incubated at 25°C.
Briefly, 4 μL of cell lysate prepared from luciferase assay was transferred to a clear 96-well plate.
A 50 µl aliquot was transferred to a clear bottom microtiter plate (Greiner) for analysis of whole cell fluorescence.
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