title: GSM7830939: TNIL_RNA_2; Homo sapiens; RNA-Seq
library_construction_protocol: After receiving written consent, myometrial biopsies from healthy women undergoing term-not-in-labor elective Caesarean sections (TNIL, n = 6) or spontaneous, non-induced term labor (LAB, n = 6) were collected. Tissues were crushed to a fine powder on dry ice and total RNA was extracted using Trizol and further treated with DNAseI for gDNA elimination. RNA was quality assessed and quantified using BioAnalyzer. RNA samples were checked on the Agilent Bioanalyzer 2100 RNA Nano chip for RNA integrity (RIN > 7) and quantified using the Qubit RNA HS Assay. RNA library preparation was performed following the Illumina® Stranded Total RNA Prep, Ligation with Ribo-Zero Plus protocol to generate stranded data. In brief, 500 ng of total RNA was used as the input material for rRNA depletion. RNA was first hybridized with DNA probes complementary to rRNA targets which direct RNase H to digest the rRNA. Any remaining DNA probes were digested with DNase I, followed by bead purification to recover remaining intact RNA. Remaining RNA was fragmented to 200-300-bases range for 2 minutes at 94oC and converted to double stranded cDNA, which proceeded to library preparation and dual-index Illumina adapters were added through PCR. Final libraries were validated on the Fragment Analyzer using the High Sensitivity NGS Kit or on the Bioanalyzer 2100 DNA High Sensitivity chip (Agilent Technologies) to check for size and absence of primer dimers, and quantified by qPCR using Kapa Library Quantification Illumina/ABI Prism Kit protocol (KAPA Biosystems). Validated libraries were pooled in equimolar quantities and sequenced on the IlluminaNovaseqplatform following Illumina's recommended protocol to generate paired-end reads of 150-bases in length.