Project description:Crude extracellular vesicles (EVs) from eight healthy volunteers were separated into 6 fractions based on their densities by using the iodixanol-based density gradient centrifugation method. To determine the distribution of miRNAs among these fractions, quantities of 93 miRNAs were quantified by the TaqMan real time PCR method using the BioMark HD system (Fluidigm) equipped with 96.96 dynamic array (Fluidigm). Six samples were fractionated from a crude EVs by density gradient centrifugation. Total of 48 samples were prepared from 8 healthy volunteers. Technical replicate of 4 gave 8 x 6 x 4 x 93 = 17,856 data. As control Tris-HCl EDTA buffer (TE) was used.
Project description:Gradient fractions of RNAi of XAC1 (Tb927.7.2780) in Trypanosoma brucei bloodstream forms. RNAi was induced using tetracycline and cell extracts were fractionated into polysomal and monosome-non-ribosome-associated fractions.
Project description:RNA-Seq of cellular and extracellular samples from DKO-1 and Gli36 cell lines. The distribution of extracellular RNA between microvesicles, and crude small extracellular vesicular vesicles fractionated based on flotation density in ioxdixanol is distinct between the different extracellular compartments.
Project description:Project for the study of spatial and temporal alterations in organelle proteins during HCMV infection. Two datasets are included, one for label-free quantification and one for TMT quantification. Samples were collected at 5 time points of infection (24, 48, 72, 96, and 120 hours) and subject to organelle fractionation. One non-infected sample was included for the label-free analysis, and 5 non-infected samples (one per time point) were included for the TMT dataset. The nucleus and cytosolic fractions were removed by differential centrifugation, leaving a crude organelle fraction. The crude organelle fraction was further fractionated by density gradient ultracentrifugation into sicx fractions.
Project description:To identify proteins present in the milked venom of the sea anemone Stichodactyla haddoni we used a bottom-up proteomics approach to analyze the digested 2DE gel spots, RP-HPLC fractions, as well as crude desalted venom.
Project description:piRNA-deficient Maelstrom (Mael) null mice are characterized by a strong upregulation of LINE-1 (L1) retrotransposon in meiotic spermatocytes. This defect turns out in the accumulation of L1 RNA and ORF1p in their cytoplasm and the formation of prominent ribonucleoprotein aggregates. We used 3-months-old Mael-/- male mice to characterize the RNA present in those ORF1p aggregates. To favor the isolation of complexed versus free ORF1p protein, we first fractionated Mael-/- testis extracts (that we refer to as TOTAL) by sucrose gradient ultracentrifugation, in the presence of EDTA. We then pooled the sucrose fractions where ORF1p macromolecular complexes sediment (fractions 5-8) and used this pool as the INPUT for an anti-ORF1p co-immunoprecipitation (IP) followed by RNA-seq.
Project description:Triton X-100 soluble crude extract from BCECs displaying limited blood-brain barrier functions was fractionated into 5 fractions (F0, F25, F50, F75) with increasing concentrations of acetonitrile (0%, 25%, 50%, 75%). This is the merging of triplicate dataset of subfraction # F0.
Project description:The sequence of gene regulatory events that drive neonatal germ cell development in the mammalian testis is not yet clear. We assessed changes in mRNA utilization in the neonatal testis at 1 and 4 dpp, times when the testis contains quiescent gonocytes (1 dpp) and proliferating spermatogonia (4 dpp). There are not thought to be major changes in the nature or number of somatic cells over that interval. We used microarrays to detail the global expression levels of mRNA distribution between non-translating mRNAs and efficiently translating mRNAs during testis development at 1 dpp and 4 dpp Extracts of mouse testes from 1 dpp and 4 dpp CD-1 mice were fractionated over a sucrose gradient in triplicate. Gradients were fractionated using an ISCO gradient fractionation system equipped with a UA-6 detector. The position of the 80S peak defined the boundary between translating and non-translating mRNAs. Pooled non-polysomal (non-translating; fractions 1-3) and heavy polysomal (efficiently translating, fractions 7-13) RNAs were isolated from sucrose gradient fractions for hybridization on Affymetrix 420 2.0 microarrays.