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HUVECs (PD 20) were treated with 75 μ m tert-butyl hydroperoxide (tBHP) in growth medium for one hour per day on 5 consecutive days as described by Dumont et al. (2000).
To evaluate the stress-tolerance of transgenic Arabidopsis seedlings, seeds were germinated and grown on 1/2 MS medium for one week, and the seedlings were transferred to soil to grow for three weeks before treatments.
For extreme temperature stress, strains were grown on rice flour medium for one week and then placed at -80°C: half the plates for 1 h and the other half for 24 h.
For mycelium fragmentation stress, strains were grown on rice flour medium for one week and the surface of the medium was watered and scrubbed, and the harvested mycelium was placed in tubes.
In the present study, two-week-old mature embryo-derived calli were cultured on fresh MSE2 medium for one week prior to bombardment resulting in a high transformation efficiency (up to 10%%) as depicted in terms of transgenic plantlets regenerated per hundred resistant calli.
They were transferred to glass tubes on a semi-solid DEV3 medium for one month under a light intensity of 60 μmol m-2 s-1 and a 12 h light/dark photoperiod for embryo conversion into plants.
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The medium for one-step fermentation of raw inulin extract from Jerusalem artichoke tubers by Paenibacillus polymyxa ZJ-9 to produce R,R-2,3-butanediol (R,R-2,3-BD) was developed.
(Optional: allow to soak overnight in milk, to rid any remaining blood). Slice into 1/2 inch thick pieces and sear in a cast iron pan over medium heat for one minute on each side.
Beat the sugar and canola oil in the bowl of an electric mixer on medium speed for one minute.
Transformed cells were cultured on S.O.C medium (Invitrogen) for one hour in a shaking incubator at 37C and then incubated overnight at 37C on LB-medium supplemented with 50 μg/ml Ampicillin and 50 μl of X-gal (40 mg/ml).
Seeds were surface sterilized by the vapour phase method (Clough and Bent, 1998) before being plated on Hoagland medium deficient for one of the four micronutrients Fe, Zn, Cu or Mn, and supplemented with 1% (w/v) sucrose and 0.5% (w/v) Fe-free agar (EDTA (ethylene diamine tetraacetic acid -washed, see Supplementacid -washed detailsee
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