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Sodium salicylate was resuspended at a stock concentration of 100 mM and diluted 1 1000 in media for experiments.
Chloramphenicol (35 μg ml−1) and ampicillin (100 μg ml−1) were added to culture media for experiments involving pSB1C3 and pSB4A5 plasmid vectors, where appropriate.
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In addition, B. subtilis was grown in competence media for transformation experiments as described [31] and MSgg medium for complex colony experiments [36].
In addition, sodium phosphate dodecahydrate was added to the growth media for these experiments, which acts as a buffer for the shorter chain fatty acids.
After three washes in KB solution, myocytes were resuspended in KB solution or in culture media for further experiments.
(Because the ilvC and ilvD gene products are also involved in valine biosynthesis [9], the selective media for these experiments was supplemented with valine).
Unless otherwise noted, media for cell experiments consisted of high-glucose bicarbonate-buffered DMEM containing L-glutamine and sodium pyruvate, supplemented with 1% antibiotics-antimyotics, 1% non-essential amino acids, and 500 µg/mL zeocin.
The final quantity of solvent did not exceed 0.1%% of culture media for all experiments.
iPS cells were induced by transduction with four Yamanaka factors using standard protocol (Okita et al., 2007), with slight modification on induction media for some experiments.
All drugs were prepared as 20 mM stock solutions in DMSO and extemporaneously diluted in media for the experiments conducted on cells.
After 6 8 h incubation at 55°C with agitation, the grown seed culture was inoculated (5%, v/v) into appropriate media for optimization experiments including temperature, medium initial pH, DO, carbon source, and nitrogen source.
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