GSM5194382: GFP_Z_2; Nicotiana benthamiana; ncRNA-Seq
Total RNA was harvested using Trizol reagent and was cut by page electrophoresis to separate 18-30 nt small RNA. (1) 3' end adaptor ligation: Ligate the 5-adenylated and 3-blocked adaptor to the 3' end of the small RNA fragment. (2) Add Unique molecular identifiers (UMI) labeled Primer.  (3) Unligated adaptors digestion.  (4) 5' end adaptor ligation.  (5) First strand synthesis with Unique molecular identifiers (UMI) labeled Primer.  (6) Second strand synthesis.  (7) Fragment selection. Page electrophoresis was used to separate PCR products in the range of 100-120 bp.   (8) Library quantitative and pooling cyclization.  (9) Library QC.