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The Ahlburg wild type strain and the deletion mutants AΔ ctb2-1were selected for transformation with DsRed resulting in more than 20 DsRed fluorescent transformants of each strain.
The best transformants of each of the gene combinations were chosen.
Thirty-six positransformantsmants of each peptide were screened by inhibition zone against S. aureus ATCC25923.
Plasmid DNA was sequenced to identify transformants of each allele.
The average and standard deviation of 8 fluctuation tests performed with 4 independent transformants of each strain are shown.
Thirty T1 double transformants of each combination were harvested and subjected to HygR and PPTR selection again to obtain single copy T2 transformants.
Similar(38)
Typically we obtained hundreds of transformants on each plate and the majority of those analyzed had the correct digestion pattern (Table 2).
According to the diameter of inhibition zone, one transformant of each peptide was selected to analyze the expression condition by Tricine-SDS PAGE.
From a pool of 75000 transformants for each block of three Cys, we identified multiple mutants that preserved drug resistance and yeast mating activity.
Eighteen of these transformants, representative of each class of promoter strength (weak, medium and strong) were precisely ranked according to the survival rate they conferred in the presence of 100 μg.ml-1 neomycin.
RNA was extracted, using RNeasy Mini Kit from Qiagen, from selected transformants representative of each strength class and grown for 48h on the surface of cellophane disks laid on solid HT medium containing only 50 μg.ml−1 thiostrepton.
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