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S37P-hTERT cells were impaired in their motility both in normal growth medium and particularly when culturing cells in poor medium (medium lacking either serum or l-glutamine).
A 50 μl volume of the suspension, as well as 10×, 100×and 1000×dilutions were dropped on solid SC medium lacking either leucine, uracil and tryptophan (growth control) or histidine, leucine, uracil and tryptophan (reporter gene assay).
At time points 0 h, 4 h and 24 h, 100 μl samples were serially diluted and plated on medium lacking either methionine (MAT α ), lysine (MAT a) or methionine and lysine (diploids).
Addition of NaH2PO4(N) instead of K to medium did not promote G. Slight G stimulation occurred at 16.6 micrograms/ml mixed aflatoxins (MA) in medium lacking either K or N but low G inhibitions were observed with K or N. The MA at 33.3 micrograms/ml reduced G 2.5% in K's of N's absence and 26 or 17% in their presence.
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Cells were pelleted and resupended in 1 ml NP-buffer (0.15 M NaCl, 10 mM Bicine pH 8.35), pelleted a second time and resuspended in 200 µl NP-buffer and plated on to SD medium (+2% agar) lacking either leucine (prey) or tryptophane (bait).
The cells were cultured on synthetic complete medium lacking uracil and tryptophan, either with histidine (SC-UW) or without histidine (SC-UWLH).
Wild-type strain (FY3027 h+) was plated either onto medium lacking arginine (-Arg) and then onto medium lacking uracil (-Ura) or directly on to double selection medium lacking both arginine and uracil (-Arg, -Ura) and mutagenised by UV (3 5 mJ, 50 80% killing).
Yeast was grown at 30°C in either standard YPD medium or minimal medium lacking leucine (leu) to select for appropriate plasmids.
Saos-2 and U2OS cells were also incubated in either Earle's Balanced Salt Solution (EBSS, a medium lacking amino acids and serum) or regular medium lacking serum or glucose.
The cells were then transfected for 4 hours with 3 µg of either pcDNA-GFP or pcDNA-β-gal DNA in fresh complete medium lacking the inhibitor.
For fluorescence microscopy, yeast cells were grown to an OD=0.6 in either YPD or, when selecting for Pkh expression plasmids, in synthetic medium lacking leucine.
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