Project description:The aim of this study was to develop a suitable method to preserve fecal samples for metaproteomics analyses when flash-freezing is not an option. Fecal samples were collected from conventional adult C57BL/6 mice and combined into a fecal master mix. The fecal master mix was then split into 48 subsamples that were subjected to different preservation treatments. The following six preservation methods were tested: flash-freezing in liquid nitrogen followed by storage at -80°C, immersion in RNAlater® and storage at room temperature, immersion in RNAlater® and immediate storage at -80°C, immersion in 95% ethanol and storage at room temperature, immersion in a RNAlater-like buffer “NAP buffer” and storage at room temperature, and immersion in an autoclaved RNAlater-like buffer “Autoclaved NAP buffer” and storage at room temperature. Proteins were extracted from the samples after being stored for 1 and 4 weeks. There were 4 replicates per treatment and time-point. Samples were analyzed by LC-MS/MS and the data were analyzed with Proteome Discoverer against a large database of mouse microbiota protein sequences.
Project description:Along with lipidomic and metabolomic analyses, we analysed the effect of short-term heat stress on Nicotiana tabacum pollen tubes. Tubes were either grown for 3 hours at room temperature, for 6 hours at room temperature or for 3 hours at room temperature and then 37 °C for another 3 hours.
Project description:Purpose: We investigated the transcriptomic change in brown fat of young and old mice (wild type) through high-throughput RNA-sequencing (RNA-Seq) analysis when the mice were exposed to cold room or room temperatur. Methods: We prepared 10 of young (3 months) mice and 9 of old (24 months) mice, and kept them in cold room (4°c) or room temperature (24°c) for 24 hours. Then, we sacrified mice and extracted RNA from brown fat tissue (BAT) for RNA-seq experiment. Results: BAT of Young mice showed increased carbohydrate metabolism and glycolytic flux during cold exposure Conclusions: The thermogenesis function of BAT is accelerated on cold exposure.
2020-07-31 | GSE135391 | GEO
Project description:A study on storabilities of Citrus grandis var.shatinyu under room temperature
Project description:Comparative transcriptome analysis of Solenopsis japonica (Hymenoptera: Formicidae) under low temperature and normal room temperature
Project description:The plant leaves were incubated (twice) at room temperature in NaCl solution (1M, 5 mL) for 15 min each, in methanol (5 mL), then in methanol-chloroform (1:1, 5 mL) twice for 30 min, and finally washed with methanol (5 mL). After each incubation, the supernatant was discarded.
The residue was dried in the open air and submitted to alkaline hydrolysis for 18 h at room temperature and in absence of light using NaOH (1M, 50 mL g-1). The alkaline extract was filtered and acidified with HCl until pH 3 and filtered again. The extract was submitted to the same process of cleanup with SPE described above, but the column was washed with ultrapure water and methanol 5% before elution using 10 mL of methanol.