RNA seq of H2O, glucose, mannose or galactose-treated Batrachochytrium salamandrivorans zoospores
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https://www.ncbi.nlm.nih.gov/sra/SRP291769
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We report RNAseq data from B. salamandrivorans zoospores treated with different carbohydrates (glucose, mannose or galactose, 50 mM) or H2O as a control, for 1 hour. We found a selective upregulation of putative B. salamandrivorans virulence factors during initial contact with carbohydrates and galactose specifically. Overall design: Total RNA was isolated from B. salamandrivorans zoospores treated with different carbohydrates. Therefore, zoospores were harvested from 175 cm2 cell culture flasks by replacing the TGhL broth with distilled water, which was filtered using a sterile mesh filter with pore size 10 µm. Six independent pools containing 4 x 10e7 zoospores were obtained (biological replicates). Per pool, the spores were divided in 4 eppendorfs (10e7 zoospores/eppendorf) which were treated for 1 hour at 15°C with H2O (control), 50 mM (a-D-galactose), 50 mM (a-D-glucose) or 50 mM (a-D-mannose). After 1 hour, the zoospores were centrifuged for 5 min at 3000 rpm at 15°C to remove the supernatant, after which RNA was extracted using the RNeasy mini kit (Qiagen). The RNA was treated with Turbo⢠DNase, following the manufacturer's instructions. RNA degradation and contamination was monitored on 1% agarose gels. The RNA purity was checked using the NanoPhotometer® spectrophotometer . Finally, the RNA integrity and quantitation were assessed using the RNA Nano 6000 assay kit of the Bioanalyzer 2100 system.
创建时间:
2021-10-15



