Project description:Here, we compile valuable insights gathered over years of generating Ribo-seq datasets from different plant species and experimental setups. We tested the effects of variable ribonuclease treatments for the generation of ribosome protected fragments (RPFs). We tested rRNA depletion strategies designed specifically for Arabidopsis and Tobacco. We also compare ligation-free to ligation-based library preparation strategies for generating Ribo-seq libraries.
Project description:Input mRNAs and ribosome protected fragments (RPFs) from wildtype and Mettl5 knockout (KO) B16 cells were analyzed by TruSeq Ribo Profile (Illumina) library preparation and high throughput sequencing
Project description:Input mRNAs and ribosome protected fragments (RPFs) from wildtype and Mettl5 knockout (KO) mESC cells were analyzed by TruSeq Ribo Profile (Illumina) library preparation and high throughput sequencing
Project description:Input mRNAs and ribosome protected fragments (RPFs) from Heterozygout (Control) and Dis3l2 knockout (KO) mESCs cultures were analyzed by TruSeq Ribo Profile (Illumina) library preparation and high throughput sequencing
Project description:To identify RDR1/2/6 dependent small RNA loci of transcribed regions under 2hr drought stress- and untreated conditions, expression profiles of small RNA between rdr1/2/6 and wild-type using hi-seq 2000. rdr1/2/6 and wild-type plants (Arabidopsis thaliana ecotype Columbia) were grown on MS medium for 2weeks. Then, their plants were transfered to dry plastic dish for 2 hours. small RNA fraction was s extracted from the sample using Ambion mirVana miRNA Isolation Kit. small RNA labrary was constructed using TruSeq Small RNA Library Preparation Kits and applied to hi-seq 2000.
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 generated 38-bp Illumina reads from messenger RNA libraries from mites transferred from their preferred laboratory host, bean (Phaseolus vulgaris cv. California Red Kidney), to one of three hosts: bean, Arabidopsis thaliana (Bla-2 accession) and the tomato (Solanum lycopersicum; genotype Heinz 1706). Larvae were carefully collected from bean plants and transferred to the treatment plant. Mites were reared on these plants for ~24 hours, after which mites were collected for mRNA library preparation. Samples were a mix of males and females. The goal of the study was to identify genes that may underlie the ability of mites to be herbivores on different host plants. Mites were transferred from host bean plants to two non-preferred hosts (Arabidopsis thaliana and tomato). RNA was then collected, and RNA-seq was performed on the Illumina platform. For each of three host plants, three biological replicates were generated.
Project description:We used Ribo-seq (Ribosome profiling) combining with RNA-seq to explore the translational landscape of Arabidopsis Col-0 seedling. We generated 6 biological replicates of RNA-seq and Ribo-seq data for Arabidopsis Col-0 seedling. 3 of the replicates were collected after 20 minutes of 0.1% DMSO treatment and the other 3 samples were collected after 60 minutes of DMSO treatmeant. The resulting RNA-seq and Ribo-seq files were used to discover translated up-stream ORFs (uORFs) and analyze the translation efficiency of uORF-containing genes in Arabidopsis.