Project description:To study the early events of priming response leading to changes in gene expression in Arabidopsis thaliana Col0 wild type plant on infiltration with the potent elicitor molecule Diffusible Signal Factor (DSF). In the present experimentation we have employed total RNA microarray expression profiling as a discovery platform to identify differentially expressing genes that are upregulated and down regulated in the various pathways involved in priming the defense responses by the elicitor molecule DSF on infiltration in the host plant Arabidopsis thaliana. This would help understand the possible defense pathways elicited by the elicitor molecule during interaction. The rosette leaves of Arabidopsis thaliana Col0 plants of 4 weeks old were infiltrated with 100uM DSF and the total RNA samples at 4hpi, 8hpi and 16hpi were extracted along with methanol treatment serving as a control in two biological replicates 1A and 2A. Further, these RNA samples were checked for the quality and subjected to microarray analysis. The differentially expressing candidate genes were identified specific to DSF treatment and were quantified by real-time PCR post DSF treatments.
Project description:We investigated lanthanide (Ln) utilization in Beijerinckiaceae bacterium RH AL1, which requires light Lns (La–Nd) for methanol oxidation. The strain was grown with methanol as the carbon and energy source and with Ln sources including natural minerals (apatite, bastnäsite, gadolinite, loparite, monazite, xenotime), an alloy (ferrocerium), and synthetic compounds (La₂O₃, Nd₂O₃, LaPO₄). We did two rounds of incubations and cultures were grown to mid-exponential phase. Total RNA was extracted and mRNA enriched by means of subtractive hybridization, before samples were subjected to sequencing library preparation for downstream Illumina high-throughbut sequencing. Pre-processed and trimmed data was subsequently used for differential gene expression analysis.
Project description:Untargeted multi-omics analysis of plasma is an emerging tool for the identification of novel biomarkers for evaluating disease prognosis and for a better understanding of molecular mechanisms underlying human disease. The successful application of metabolomic and pro-teomic approaches relies on reproducibly quantifying a wide range of metabolites and proteins. Herein, we report the results of untargeted metabolomic and proteomic analyses from blood plasma samples following analyte extraction by two frequently used solvent systems: chloro-form/methanol and methanol-only. Whole blood samples were collected from participants (n=6) at University Hospital Sharjah (UHS) hospital, then plasma was separated and extracted by two methods i. methanol precipitation and, ii. 4:3 methanol:chloroform extraction. The coverage and reproducibility of the two methods were assessed by ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). The study revealed that metabolite extraction by methanol-only showed greater reproducibility for both metabolomic and proteomic quantifications than did methanol/chloroform, while yielding similar peptide coverage. However, coverage of extracted metabolites was higher with the methanol/chloroform precipitation.
Project description:We examined changes in steady-state transcript level in leaves of Arabidopsis plants subjected to salinity, heat stress and their combination by a transcriptome analysis of leaves.