Small RNA seq of Piper n., stem-repeat2
3 g of total RNA per sample was used. NEB 3' SR Adaptor was first ligated to the 3 end. SR RT Primer hybridized to the excess of 3' SR Adaptor and transformed the single-stranded DNA adaptor into a double-stranded DNA. 5 ends adapters were ligated to the 5 ends of small RNA fragments. A RT reaction was then performed to generate cDNA. PCR amplification was performed. PCR products were purified, recovered and dissolved in elution buffer.