small RNA-seq of 'changchun': vegetative stage
Total RNA was extracted from the fresh plant tissues which were collected above using improved CTAB-PVP method, The small RNAs with certain size (18-30 nt) were separated using polyacryamide gel electrophoresis (PAGE). First strand cDNA was synthesized after the selected fragments were successively subjected to 3 adapter ligation, RT primer hybridization and 5 adapter ligation. Subsequently, PCR enrichment, size selection by PAGE, heat separation and ssDNA cyclization were performed.