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Fig. 1 A representative screening plate used for selection of UV mutants.
About 300 highly mucoid colonies were picked up from the screening plate, and then transferred into the basal fermentation medium.
Listed below are steps followed for each generation of mutagenesis with slight variations based on the screening plate used in first step of selection.
The display was designed so that it could simultaneously display the images of a 96 well screening plate, corresponding to 8 pyridinium or quinolinium building blocks conjugated to 10 aldehyde building blocks, as well as images from the outer two columns of control wells, labels for each image and chemical structure of the probes (Figure 1C).
Next, four IRPs were then transferred (10 µl of virus/well) into each screening plate in triplicate by use of a Biomek® FX Laboratory Automation Workstation (Beckman Coulter, Fullerton, CA).
Inter-plate variability in period length (25.82±0.97 h, n = 4) and amplitude (106±76, n = 4) was addressed by running scrambled siRNA transfected wells on every screening plate.
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Two such screening plates are shown in Fig. 2.
Next day, mycelia were separated from spores using cotton wool, broken down using glass beads and plated under different dilutions on various screening plates.
The initial growth on the screening plates might be due to the presence of intracellular storage compounds accumulated during the revitalization stage in rich medium.
a Gram's iodine staining, showing the distinct zones of clearance of the alginate lyase-excreting strains (one of the screening plates for result exhibition).
Aliquots (100 μl) of diluted sample were spread on screening plates consisted of 0.5% sodium alginate, 0.5% ammonium sulfate, 0.2% dipotassium phosphate, 0.1% magnesium sulfate, and 2% agar (ALG plates) (pH 7.2 7.4).
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