title: GSM5563754: Mouse seminal vesicle tissue transcriptomics data - Control vehicle (PBS treated) 1; Mus musculus; RNA-Seq
library_construction_protocol: Seminal vesicle samples were homogenised using the FastPrep-24 5G homogeniser (MP Biomedicals, Irvine, CA) with the Cool Prep Adaptor (2 × 1min, 4.0m/s, at 4 °C) in Trizol reagent (ThermoFisher Scientific, Waltham, MA). Total RNA was extracted using the Trizol RNA method and DNase treated using RQ1 RNase free DNase (Promega, Madison, WI), following the manufacturer's instructions. RNA was quantified using a Nano-Drop Lite Spectrophotometer (ThermoFisher Scientific) and RNA integrity was assessed using an Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA). RNA with a RIN > 7 was used in this study. RNA sequencing was completed at BGI (Shenzen, China). RNA-sequencing analysis was performed at BGI Genomics (Shenzhen, China) using DNBseq technology. Poly-A libraries were constructed by initially purifying poly-A mRNAs using poly-T oligo attached magnetic beads. Poly-A mRNAs were fragmented using divalent cations and the cleaved RNA was processed through first strand cDNA synthesis using reverse transcriptase and random primers, followed by second strand cDNA synthesis using polymerase I and RNase H. Second strand cDNA was then purified, and DNA nanoballs were generated with the libraries by rolling circle replication and samples were loaded into patterned nanoarrays to generate 100 bp paired end reads through the DNBseq platform. For each library, a minimum of 67 million paired end raw reads were produced.