A recent study focused on fission yeast strains and growth conditions, specifically S. japonicus and S. pombe strains. The work utilized standard fission yeast media and culture methods, including a modified YNB medium to support growth under strictly anaerobic conditions. Liquid cultures were grown in shaking incubators at 30°C using YES or modified YNB medium. Anaerobic cultures were established within an InvivO2 workstation. Lipidomics analyses were conducted for S. pombe in anoxic conditions. Mating of S. japonicus and S. pombe was induced on solid medium, and spores were germinated for further analysis.
Various materials were used in cell biological and biochemical experiments, including chemicals like D-glucose, agar, and different reagents for experimental setups. Molecular genetics manipulations were performed using PCR or plasmid-based recombination. Serial dilution assays were conducted for S. japonicus and S. pombe cells to assess growth under different conditions. Genetic crosses between strains were performed to study spore lethality and mutant viability.
Microscopy and imaging analyses were conducted to visualize cellular structures and substances such as sterols. Cell growth, colony-forming units, Western blotting, and RT-qPCR experiments were conducted to assess cellular properties. Triterpenoid detection and lipidomic analyses were performed using GC-MS and ESI-MS techniques. Biophysical experiments, such as vesicle preparations, water permeability measurements, and fluorescence anisotropy studies, were also carried out.
The study included statistical analyses, replicates, and comprehensive reporting. The experimental processes were meticulously detailed, and a range of experiments was conducted to explore various aspects of fission yeast biology. The results provide valuable insights into the growth, genetics, physiology, and lipid composition of fission yeast strains. The findings contribute to the understanding of cellular processes in these organisms and offer potential applications in diverse research areas.
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