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If the growth of a mutant on the plate containing certain reagent was 2-spot lesser than that on YES plate (25 fold reduction in viability), this mutant was designated as sensitive.
An acetone-soaked filter paper (Whatman #3, 1003-090) was placed on a glass Petri dish and inverted above the YES plate to expose the patches to acetone vapor for 15 min. These were then incubated at 31° for 3 d before being photographed.
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Serial dilutions of the wild-type and respective mutants were plated on YES, and YES plates containing FOA. DAPI staining were used to determine the percentage of lagging chromosomes as described previously (Gregan et al. 2007).
Cells were grown to mid-log phase, serially diluted five-fold and spotted onto YES plates or YES plates were irradiated with the indicated doses of UV irradiation using a Stratalinker (Stratagene) or YES plates containing 10 or 12 μg/ml thiabendazole (TBZ), 5 mM hydroxyurea (HU), 7 μM camptothecin (CPT), 0.005% methyl methanesulfonate (MMS) and incubated for 2-3 dats at 32°C.
Random spores were germinated on YES plates to form colonies.
YES plates were replica plated onto EMM plates lacking histidine, lysine, adenine, or histidine and lysine.
Transformants were selected on YES plates containing 0.1 mg/mL nourseothricin.
Transformants were selected on YES plates containing 0.5 μg/mL aureobasidin A (Takara).
YES plates were incubated at 32° for 3 d and PMG-ura-ade plates for 5 d before colonies were counted.
When cultures reached a stable maximal density, cells were harvested, serially diluted, and plated on YES plates.
Following heat treatment, spores were transferred on YES plates and allowed to germinate for 2 days at 32°C.
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