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Concentrated stock cultures of B. breve B632 were supplemented with glycerol (10%, v/v), enumerated onto MRS-agar plates, and stored at −80°C until an appropriate volume was thawed and used for bioreactor inoculation.
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Colonies were enumerated, isolated onto plates containing trypticase soy agar with 5% sheep blood, and incubated for 24 hrs.
The dilution representing the MIC and two of the more and less concentrated dilutions were plated out onto Trypticase soy agar plates and enumerated to determine viable CFU/ml (31).
The bacteria were enumerated on TSA.
E. coli O157 H7 in cultures was serially diluted and then enumerated by direct plating onto tryptic soy agar supplemented with 50 mg/L nalidixic acid (TSA-nal; Dalynn Biologicals, Calgary, AB, Canada) after incubation for 18 h at 37°C.
Bacteria were enumerated by plating serial dilutions onto XLT-4 plates (Beckton Dickinson and Company, Franklin Lakes, NJ).
Bacteria were enumerated after plating a dilution series onto LB agar.
Caco-2 cells were then lysed with 0.1% Triton-X 100 in PBS and adherent bacteria enumerated by viable counts following plating onto Columbia agar with 5% horse blood and grown at 42°C for 36 h.
To exclude effects on bacterial viability, blood was plated onto blood agar plates and CFUs enumerated, after overnight incubation (37°C/5% CO2).
Samples were enumerated by a 10-fold dilution series, plating each dilution onto TSA, and counting the colony forming units (CFU) after sufficient growth at 37°C overnight.
Bacterial eluate solutions were enumerated by a 10-fold dilution series, plating each dilution onto TSA, and counting the colony forming units (CFU) after sufficient growth at 37°C overnight.
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