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Diazoxide capsules were administered with 240 mL (±5 mL) of room temperature, potable water after at least 10 hours of overnight fasting.
Group W received 200 ml of heated carrier fluids for 20 minutes prior to 2 mg/kg propofol injection; group L received 200 ml of heated carrier fluids for 20 minutes prior to lidocaine pretreatment and 2 mg/kg propofol injection: and group C (control group) received 200 ml of room temperature fluids prior to 2 mg/kg propofol injection.
A longitudinal view of the fossa ovalis was obtained to evaluate right-to-left shunting by injecting 9.5 mL of sterile-modified fluid gelatine solution (Plasmion [Fresenius-Kabi, Sevres, France] or Gelofusine 4% [B-Braun Medical, Boulogne-Billancourt, France]) aerated with 0.5 mL of room air via two syringes connected with a three-way stopcock, as previously described [2,11].
The supernatant was removed, and the pellet was resuspended in 30 ml of room temperature stop buffer.
Tissue was immediately placed in 4 mL of room temperature RNAlater for 2 minutes with mixing, transported on ice, held overnight at 4°C, and placed at −20°C until total RNA isolation.
24 hours after addition of FUDR to the plates, worms were transferred using a worm pick to a well of a six well cell-culture plate containing 3 ml of room temperature S-basal/cholesterol/antibiotics solution+FUDR (100 mg/L).
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By using a dissolution insert that fits to the new design, it was possible to obtain about 120 ml of room-temperature hyperpolarized solution.
Male 12-week-old Long Evans rats, either WT or KO, were deeply anaesthetized with isoflurane and transcardially perfused with 300 ml of room-temperature (25°C) saline.
Male 12-week-old C57BL/6J mice, either WT or LRRK2-KO [ 10], were deeply anaesthetized with isoflurane and transcardially perfused with 50 ml of room-temperature saline under isoflurane anaesthesia.
From each tube, 400 μl of the aqueous phase was transferred to a fresh 1.5-ml tube, 400 μl of 100% isopropanol added, mixed by vortex and incubated at room temperature for 10 min. The tubes were then centrifuged at 7,500 × g for 10 min at 4°C and the supernatant removed by pipette and 1 ml of room-temperature 75% ethanol added.
Incremental 1 ml volumes of room air were injected via the tracheal cannula, and the pressure attained 3 seconds after each injection was measured, until a total volume of 5 ml was injected.
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