microRNA sequencing of human breast cancer
For small RNA library preparation, 100 ng of RNA was subjected to 3' ligation followed by 5' ligation and reverse transcription. Subsequently, QIAseq miRNA NGS (QMN) bead preparation, cDNA cleanup, and finally library amplification was carried out using tube indices (QIAseq miRNA NGS 96 Index IL kit, QIAGEN, Hilden, Germany). The yield of cDNA libraries was quantified using Qubit dsDNA HS assay kit (Invitrogen) and size distribution of the cDNA libraries were determined using the Agilent 2100 Bioanalyzer DNA1000 chip (Agilent Technologies). Libraries were subsequently pooled and were subjected to sequencing using Illumina HiSeq 4077