title: miRNA-Seq of Human granulosa cells: adult female
design: 1g of total RNA was used as the input material for RNA sample preparation using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA). For small RNA sequencing, briefly, 3 and 5 adaptors were ligated to 3 and 5 end of small RNA, respectively. Then the first strand cDNA was synthesized after hybridization with reverse transcription primer. A double-stranded cDNA library was generated through PCR enrichment. After purification and size selection, libraries with insertions between 18~40 bp were ready for sequencing on Illumina sequencing with SE50. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Qualified libraries were pooled and sequenced by the Illumina NovaSeq 6000. Six biological replicates were performed for each sample.