GSM5495165: Fetal chondrocytes; Homo sapiens; miRNA-Seq
RNA was isolated using miRNeasy Micro Kit (Qiagen) according to manufacturer's protocol. Briefly, samples were lysed by QIAzol lysis reagent followed by addition of chloroform and centrifugation to separate the solution into phases. The upper aqueous phase was extracted, ethanol was added, and samples were loaded into RNeasy MinElute spin column.  Thereafter a specialized protocol was used to separate the enriched miRNA fraction. miRNA was quantified using Qubit fluorometer, and run on Agilent Bioanalyzer 2100 for quality control. Libraries were prepared using NEBNext Multiplex Small RNA Library Prep Set (Illumina) according to the manufacturer's protocol. The workflow consists of adapter ligation, cDNA synthesis, PCR enrichment, clean up and size selection. Different adapters were used for multiplexing samples in one lane. Sequencing was performed on Illumina HiSeq 2500 with single-end 50 base pair reads. NEBNext Multiplex Small RNA Library Prep Set (Illumina)