title: GSM4305308: PS16.14.04; Homo sapiens; miRNA-Seq
library_construction_protocol: Cryopreserved sperm samples were thawed, suspended in PureSperm Buffer (Nidacon), then mature sperm were enriched by centrifugation (300 g, 15 min) through a 50% PureSperm density gradient (Nidacon). Sperm were then lysed in TRIzol-LS (Thermo Fisher) reagent, supplemented with 0.2 M β-mercaptoethanol and 100 mg of nuclease-free stainless-steel beads, by homogenization on a Disruptor Genie (Scientific Industries) at 3000 rpm for 5 min. RNA, enriched for small RNA, was isolated using Qiagen's miRNeasy Mini kit according to manufacturer's instructions. Libraries were constructed using the TruSeq small RNA Library Prep Kit (Illumina) with 10 ng of small RNA according to the manufacturer's protocol. Post-PCR cleanup and size selection for products > 100 bp was performed using AMPure XP bead purification. Library size distribution and quantification was performed using on a TapeStation 4200 (Agilent) using their High Sensitivity D1000 screentape. Individually barcoded libraries were pooled to achieve ~10 million reads per sample and sequenced on an Illumina NextSeq 550 (36-bp single-end).
attributes.tag: GEO Accession
attributes.value: GSM4305308