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To verify the mating type and examine the mating abilities of the derivative strains, a mating assay was performed with the auxotrophic and G418-resistant laboratory haploid strains HR42-11T (α-type) and MCF4741 (a-type) (Fukuda et al. 2013b) (Table 1).
Mating assay was performed as described previously [25], [29].
The yeast two-hybrid mating assay was carried out as described in material and methods.
Under the above conditions, the detection sensitivity of the mating assay was >1 × 10 transconjugants/recipients.
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Images showing colony formation on SD/MSG + G418 plates in the mating assay are shown.
Using a Tukey's Honestly Significant Difference test, we found that the outcrossing rate of S. cerevisiae in mass mating assays was significantly greater than all other assay and species combinations.
The only statistically significant difference observed in these latter mating assays was opposite to that predicted by the hypothesis that males learn to avoid fruitless courtship of heterospecific females (i.e. the experienced males of the Mather, California, strain elicited a higher frequency of wing vibrations than the corresponding naïve males in the second mating assays).
Mating assays were performed on V8 medium at pH = 5 and pH = 7 [20] and MS medium, pH = 4 [21].
Mating assays were performed on SOE plates (1% sucrose, 0.5% dextrose, 0.5% fructose, 0.1% yeast extract, 0.15% peptone, 2% agar) [30].
Mating assays were carried out on V8 medium [69] (5% [vol/vol] V8 juice, 2 mM KH2PO4, 4% [wt/vol] agar).
Mass mating assays were performed once for each of all the pair-wise combinations of the twelve heterothallic strains, as well as for all combinations of the five homothallic S. cerevisiae strains.
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