Project description:Identification of new and unpredicted full length Arabidopsis genes. Examination of cRNA prepared from Arabidopsis thaliana ecotype Columbia light grown 7-day old seedlings using whole genome tiling arrays. Keywords: other
Project description:To investigate differences in plant responses to salt and ABA stimulus, differences in gene expression in Arabidopsis in response to salt and ABA were compared using an Agilent oligo microarray. Four-week-old Arabidopsis thaliana ecotype Columbia (Col-0) seedlings were treated with either 150 mM NaCl or 10 μM ABA for 6 hours; unstressed seedlings (control sample) were collected in parallel to avoid the possible effects of circadian rhythms. The results revealed that 31 genes were up regulated by both NaCl and ABA stress, and 23 genes were down-regulated by these stressors. To provide further validation of our microarray experiment data, ten genes from this signature were quantified in the same RNA samples by quantitative real-time PCR.
Project description:Comparison of TopHat alignments and assessment of spurious splice junctions for 32nt and 76nt read lengths. Total RNA from 2-week-old Arabidopsis thaliana (ecotype Columbia) seedlings grown on MS plates was isolated using RNeasy Plant Mini Kit from Qiagen. To remove any contaminating DNA, RNA was treated with DNAse. Isolation of poly (A) mRNA and preparation of cDNA library were carried out using the Illumina TrueSeq RNA kit. Sequencing (72 cycle) was done on Illumina Genome Analyzer II.
Project description:We sequenced the poly(A)+ and poly(A)- samples of the roots and shoots from 10-day-old WT seedlings grown under P+ and P- condition. The WT plant refers to Columbia ecotype Arabidopsis seedlings. Each condition has two replicates. After total RNA extraction, ribosomal RNAs were removed using RiboMinus™ Plant Kit repeated two times. The poly(A)+ and poly(A)- constituent were separated with oligo(dT) magnetic beads (Oligotex mRNA Mini Kit, QIAGEN). Using a 2-fold change and a P-value <0.05 as the cut-off for selecting the differentially expressed transcripts, we globally identified novel noncoding lncRNAs.
Project description:Identification of new and unpredicted full length Arabidopsis genes. Examination of cRNA prepared from Arabidopsis thaliana ecotype Columbia light grown 7-day old seedlings, light grown 7-day old seedlings treated at 4 degrees C for 24hrs, and etiolated 7-day old seedlings using pilot genome tiling arrays. Total RNA was isolated from seedlings with the TRIzol reagent procedure of Invitrogen, then poly(A)+ RNA was purified with Qiagen oligotex. One microgram poly(A)+ RNA was converted into ds-cDNA using T7-oligo(dT) primers. Biotin-labeled cRNA was generated by in vitro transcription reactions using ENZO labeling kit, fragmented and then 20 micrograms of fragmented cRNA were hybridized to the arrays according to Affymetrix instructions. Keywords: other
Project description:To investigate differences in plant responses to salt and ABA stimulus, differences in gene expression in Arabidopsis in response to salt and ABA were compared using an Agilent oligo microarray. Four-week-old Arabidopsis thaliana ecotype Columbia (Col-0) seedlings were treated with either 150 mM NaCl or 10 M-NM-<M ABA for 6 hours; unstressed seedlings (control sample) were collected in parallel to avoid the possible effects of circadian rhythms. The results revealed that 31 genes were up regulated by both NaCl and ABA stress, and 23 genes were down-regulated by these stressors. To provide further validation of our microarray experiment data, ten genes from this signature were quantified in the same RNA samples by quantitative real-time PCR. Differentially expression genes of Arabidopsis thaliana were measured under salt stressed, ABA stressed and normal condition for 6 hours, respectively. Three independent experiments were performed at each treatment using different plants for each experiment.
Project description:The goal of this study was to compare the transcriptional profile (RNA-seq) of Arabidopsis thaliana Columbia-0 ecotype seedlings that were treated with two different light (white light or shade) and temperature (20ºC or 28ºC) conditions.
Project description:Comparison of TopHat alignments and assessment of spurious splice junctions for 32nt and 76nt read lengths. Total RNA from 2-week-old Arabidopsis thaliana (ecotype Columbia) seedlings grown on MS plates was isolated using RNeasy Plant Mini Kit from Qiagen. To remove any contaminating DNA, RNA was treated with DNAse. Isolation of poly (A) mRNA and preparation of cDNA library were carried out using the Illumina TrueSeq RNA kit. Sequencing (72 cycle) was done on Illumina Genome Analyzer II. 2 replicates
Project description:Proteins from plant shoot and root tissues were extracted from wild-type Arabidopsis thaliana ecotype Columbia (Col-0). They were enriched on conditioned U(VI)-loaded and U(VI)-free Duolite C467 beads. The enriched proteins were identified and quantified by label-free shotgun proteomics.
