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The test enables repeated assessments (20 30 min per time point) over a school year.
A crude extract was prepared by decoction of dried root and rhizome of A. dahurica (3.0 kg) in methanol (3 L) for 3 times (120 min per time).
(A and B ) Time course imaging of 250 nM NAA treated seedlings were performed every 15 min. Image acquisition took 10 min per time point.
Then, the sections were infused in xylene (10 min per time for three times), followed by infusion in 90% ethanol (3 min), 75% ethanol (3 min), 50% ethanol (3 min) and ddH2O (3 min).
The reaction was stopped by spotting 25 µl on to a piece of P81 filter paper, which was then washed four times at 15 min per time in 1% (vol./vol)./vol
For detailed, leaves were cut into sections of 1 mm × 1 mm and fixed in 2.5 % glutaraldehyde in 0.1 mol/L phosphate buffer (PBS, pH 7.2) at 4 °C for 3 days, following by washing with 1 mol/L PBS for three times, 15 min per time.
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Then, they were decocted with water (2500 mL, 1500 mL, and 1500 mL) for 3 times, 30 mins per time, respectively.
At given time intervals of illumination (5 min per each time interval), the toluene concentration was analysed by a UV Vis spectrophotometer UV-36000, Shimadzu, Japan).
The time for respective 5 min, 30 min, 60 min and 120 min (Six rats per time point) post administration was designated to collect the cerebrospinal fluid (CSF), brain and other tissues after cardiac perfusion through left ventricle using the perfusion method described previously [38] to determinate exactly the amount of honokiol.
Wells received 10 µl of 3 mM MNNG (for a final concentration of 100 µM) at 5-min intervals over a total time course of 30 or 240 min (three replicates per time point).
The acquisition time for images with both radiotracers varied between 7 min and 10 min per view (the time required to acquire 2,000,000 2,500,000 counts per image).
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