Project description:To examine the effects of microRNAs including miR-223 on murine neutrophil function, small RNAs cloning and sequencing from neutrophils were performed. Neutrophils were isolated from a wild-type mouse and total RNAs were prepared. Small RNAs were cloned and sequenced by Solexa/Illumina genome analyzer.
Project description:Right ventricular small RNA profiles from 22 patients with Tetralogy of Fallot (TOF) and small RNA-seq profiles from the left and right ventricle (LV and RV, respectively) of 4 healthy unaffected individuals (NH) were generated. The total RNA was isolated from the 30 human heart samples using TRIzol. Small RNAs were isolated from total RNA and prepared for microRNA sequencing using Illumina Kit FG-102-1009 according to the manufacturer's protocol (Preparing Samples for Analysis of Small RNA Illumina 2007). The sequencing libraries were generated using a non-strand-specific library construction method. The purified DNA fragments were used directly for cluster generation, and 36 cycles of single-end read sequencing were performed using the Illumina Genome Analyzer. Sequencing reads were extracted from the image files using the open source Firecrest and Bustard applications (Illumina Genome Analyzer pipeline 1.3.2).
Project description:To examine the effects of microRNAs including miR-223 on murine neutrophil function, small RNAs cloning and sequencing from neutrophils were performed.
Project description:High throughput seqeuncing of small RNAs (PAGE isolated from total RNA or Argonaute immunoprecipitates) from Drosophila melanogaster using the Illumina platform. Adapter ligation requires 5' monophosphate and 3' OH. Full analysis of all libraries in this set is published (Czech B. et al. 2008), leading to the description of endogenous siRNAs in flies. Keywords: Solexa sequences
Project description:Total RNAs from the Citrus red mite at the embryo, larva, nymph and adult stages were used to construct small RNA libraries for Solexa sequencing. Several categories of sRNAs were identified, including 594 known microRNAs (miRNAs) grouped into 206 families and 31 novel miRNAs. In addition, according to bioinformatics analysis and S-Poly(T) miRNA assays, the expression level of many miRNAs varied among the developmental stages. Furthermore, the prediction of miRNAs target genes and their functional annotation indicated that miRNAs are involved in the regulation of multiple pathways in the Citrus red mite. The total RNAs from embryo, larva, nymph and adult stages of Panonychus citri were used to construct small RNA libraries for Solexa sequencing. The data from Solexa sequencer (Illumina) was compared to identify small RNAs in different stages of Panonychus citri.
Project description:Transposable elements comprise a large proportion of animal genomes. Transcripts of transposable elements are a source for the synthesis of endogenous siRNAs and piRNAs. In order to determine if small RNAs mapped to expressed Tc1-like elements are present during early Xenopus tropicalis development, we used Illumina (Solexa) to sequence small RNAs from gastrula-stage embryos. We obtained about 17 million reads that mapped perfectly to the genome. Small RNAs mapped to selected transposable elements were characterized and the expression of selected small RNAs was experimentally verified during development. This is the first deep sequencing experiment for small RNAs in the Xenopus tropicalis gastrula.
Project description:Transposable elements comprise a large proportion of animal genomes. Transcripts of transposable elements are a source for the synthesis of endogenous siRNAs and piRNAs. In order to determine if small RNAs mapped to expressed Tc1-like elements are present during early Xenopus tropicalis development, we used Illumina (Solexa) to sequence small RNAs from gastrula-stage embryos. We obtained about 17 million reads that mapped perfectly to the genome. Small RNAs mapped to selected transposable elements were characterized and the expression of selected small RNAs was experimentally verified during development. This is the first deep sequencing experiment for small RNAs in the Xenopus tropicalis gastrula. Analysis of small RNAs expressed in the Xenopus tropicalis gastrula.
Project description:The total RNA were extracted from pooled tissues of leaves and flowers from several plants of sacred lotus using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. Then small RNAs ranging in 18–30 nucleotides were size fractionated electrophoretically, isolated from the gel, ligated with the 5′ and 3′ RNA adapters. The ligated product was reverse transcribed and subsequently amplified using 10–12 PCR cycles. The purified PCR product was sequenced using Illumina Genome Analyzer II. The qualified reads were used to predict phased small interfering RNAs from Chinese sacred lotus (Nelumbo nucifera Gaertn.).