Rattus norvegicus
Protocols: EVs from rat AFSCs were isolated from cells that were treated with exosome-depleted FBS (ThermoFisher, Waltham, MA) for 18 hours by ultracentrifugation as previously described [Antounians et al, 2018] Treatment with EVs is with 10% (by volume) EVs were processed for total RNA isolation using the SeraMir Exosome RNA Purification Column kit (System Biosciences, Palo Alto, CA) according to the manufacturer's instructions. For each sample, 1μL of the final RNA eluate was used for measurement of small RNA concentration by Agilent Bioanalyzer Small RNA Assay using Bioanalyzer 2100 Expert instrument (Agilent Technologies, Santa Clara, CA). Small RNA libraries were constructed with the CleanTag Small RNA Library Preparation Kit (TriLink, San Diego, CA) according to the manufacturer's protocol. The final purified library was quantified with High Sensitivity DNA Reagents (Agilent Technologies, Santa Clara, CA) and High Sensitivity DNA Chips (Agilent Technologies, Santa Clara, CA). The libraries were pooled, and the 140bp to 300bp region was size selected on an 8% TBE gel (Invitrogen, Carlsbad, CA). The size selected library is quantified with High Sensitivity DNA 1000 Screen Tape (Agilent Technologies, Carlsbad, CA), High Sensitivity D1000 reagents (Agilent Technologies, Carlsbad, CA), and the TailorMix HT1 qPCR assay (SeqMatic, Fremont, CA).
ENA-FIRST-PUBLIC
2021-03-01T12:16:09Z
ENA-LAST-UPDATE
2020-04-11T12:25:22Z
External Id
SAMEA6663534
INSDC center name
Developmental and Stem Cell Biology Program, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children
INSDC first public
2021-03-01T12:16:09Z
INSDC last update
2020-04-11T12:25:22Z
INSDC status
public
Submitter Id
E-MTAB-8921:Sample 41
broker name
ArrayExpress
cell type
mesenchymal stromal cell
common name
Norway rat
developmental stage
not available
disease
normal
fraction
extracellular vesicle
genotype
wild type genotype
organism part
mesenchyme
sample name
E-MTAB-8921:Sample 41
scientific_name
Rattus norvegicus
sex
not available
strain
Sprague-Dawley