miRNA-Seq of yeast:WT-1
Total RNA was extracted, and small RNA fragments (20200 nt) were isolated from total RNA via gel purification. 3 and 5 RNA adapters were sequentially ligated to the ends of small RNA molecules. First-strand cDNA was synthesized by reverse transcription, followed by PCR amplification. After size selection and purification, the constructed library was quantified using Qubit and real-time PCR, and fragment size distribution was verified with a bioanalyzer before sequencing.