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A. rhizogenes strains ATCC15834 (Food Industry Development Institute, Taiwan) were grown overnight on BEP medium (beef extract and peptone) at 28°C and 180 rpm in the dark.
Then, the sample was serially diluted in sterile distilled water and followed by plating on the carbon-rich nutrient agar medium (beef extract 0.3%, peptone 0.5%, sodium chloride 0.5%, glucose 1%, and agar 2%).
Motility assays were conducted on semi-solid SM medium (beef extract at 3 g/L, peptone at 5 g/L, and 25 ml/L of 20%% glucose) with 0.5 % of agar.
Acidovorax avenae subsp avenae obtained from the Culture Collection of the Instituto Biológico (IBSBF) was grown in NA medium (beef extract 3 g/L; Peptone 5 g/L; NaCl 5 g/L) at 28°C.
Briefly, cells were prepared by growing in Muller Hinton (MH) medium (beef extract 2 g l−1, casein acid hydrolysate 17.5 g l−1, starch 1.5 g l−1, agar 17 g l−1) for 17 h at 37°C.
Medium beef should be between 135 and 145 degrees, bright pink but slightly less juicy than medium-rare beef.
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Both bacteria and yeasts were grown on nutrient agar medium (g/l): Beef extract, 3; peptone, 10; and agar, 20.
The culture was maintained on the nutrient agar medium (w/v) (beef extract 0.3%, peptone 0.5%, sodium chloride 0.8%, and agar 1.5%) and stored at 4°C.
As the selective medium contains beef and yeast extract, M9 medium was used to validate whether E. sp. CGMCC 5087 can biosynthesize 2-PE using glucose as the sole carbon source.
For sporulation supernatant preparation, 0.25 g wet weight of bacteria (corresponding to 0.5 0.7 × 109 cells) were suspended in Difco sporulating medium (bacto beef extract 3 g/l, peptone 5 g/l, NaOH 0.25 mM, MgSO4 10 mM, KCl 0.1%, MnCl2 0.1 mM, Ca(NO3 2 1 mM, FeSO4 1 mM, pH 6.8) without chloramphenicol and incubated at 35°C with shaking.
Medium rare beef, veal, and lamb steaks need to be cooked to 145ºF/62.7ºC.
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