title: GSM3384013: C1: Uncomplicated Pregnancy Control 1; Homo sapiens; miRNA-Seq
library_construction_protocol: Phlebotomy was performed by a trained health professional. In brief, 2ml of whole blood were taken from each subject and remained in an EDTA tube for 30min. Tubes were immediately thereafter centrifuged for 10min at 2000rpm. Plasma was removed and stored in 2ml Eppendorf tubes. Plasma samples were kept immediately thereafter at a -80oC storage until further processing. For total RNA extraction, 100ul of plasma were added to 1ml TRIzol (Life Technologies, Foster City, CA), treated with DNase I (Ambion, Austin, USA) and enriched for small RNAs (siRNAs, miRNAs) using the mirVana microRNA Isolation kit (Ambion, Austin, USA). RNA quantity and purity were measured using a Nanodrop spectrophotometer (ND-1000, NanoDrop Technologies, Houston, TX, USA). Total RNA of each sample was used to prepare the miRNA sequencing library, which included the following steps: 1) 3'-adapter ligation; 2) 5'-adapter ligation; 3) cDNA synthesis; 4) PCR amplification; 5) size selection of ~130-150 bp PCR amplified fragments (corresponding to ~15-35nt small RNAs). The libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 51 cycles on Illumina Nextseq per the manufacturer's instructions.
identifiers.namespace: GEO
identifiers.id: GSM3384013