GSM5598460: CVS_M_72287; Homo sapiens; miRNA-Seq
Tissue was submerged in RNAlater RNA stabilization reagent (QIAGEN) and stored at -80C until further processing. Tissue was thawed on ice and homogenized using sonication and/or needles, then total RNA was isolated using the AllPrep DNA/RNA/miRNA Universal kit (QIAGEN). The total RNA elution was used to prepare a miRNA sequencing library using the QIASeq miRNA Library Kit. A pre-adenylated DNA adapter was ligated to the 3' ends of miRNAs, followed by ligation of an RNA adapter to the 5' end. A reverse-transcription primer containing an integrated Unique Molecular Index (UMI) was used to convert the 3'/5' ligated miRNAs into cDNA. After cDNA cleanup, indexed sequencing libraries were generated via sample indexing during library amplification, followed by library cleanup. Libraries were sequenced on a NextSeq 500 (Illumina, San Diego, CA) with a 1×75 bp read length and an average sequencing depth of 10.64 million reads per sample.