title: GSM4411624: Extracellular Vesicle small RNAseq; Caenorhabditis elegans; ncRNA-Seq
library_construction_protocol: EVs were isolated from young adult N2 animals. Briefly, conditioned media from ~500,000 developmentally synchronized young adult animals are incubated in S Basal for 24 hours. The conditioned media is then processed with filtering, exclusion chromatography, and ultrafiltration. To quantify EV-associated RNA samples were processed with Total exosome protein and RNA isolation kit (Invitrogen, Carlsbad CA, USA, Cat # 4478545). We then characterized the abundance and size of the eluates on an Agilent 2200 TapeStation (Agilent, Santa Clara CA, USA; Cat # G2991AA) using high-sensitivity screen tape (Agilent, Santa Clara USA Cat # 5067-5579) along with a small RNA calibration ladder (Agilent, Santa Clara CA, USA; Cat # 5067-1550).  The vesicle samples displayed substantial small RNAs as well as 16s and 28s rRNA species at the expected sizes. To determine what RNA species are packaged into EVs we conducted RNAseq. We used a Qiagen small RNA Sample Preparation kit to prepare a cDNA library from our EV-associated RNA (Qiagen Germantown MD, USA Cat # 331502) while an Illumina MiSeq v2 kit (300 cycles) was used to prepare the sequencing library with 5' adapter sequences (San Diego USA; Cat# MS-102-2002). The sample was then submitted to single-end sequencing on an Illumina MiSeq desktop sequencer.
attributes.tag: GEO Accession
attributes.value: GSM4411624