HCT116_DROSHA_KO_d220_2_RNAseq
Total RNA in DROSHA KO HCT116 cells was isolated with RiboEX solution and separated with formamide loading buffer on 15 % urea-polyacrylamide gel at 15 W for 45 min. RNAs between 18  30 nt RNA markers were purified and 3' adaptor was ligated with T4 RNA Ligase 2, truncated (New England Biolabs, USA). The resulting RNAs were resolved on 15 % urea-acrylamide gel. 35  52 nt RNA was purified and 5' adaptor was ligated with T4 RNA Ligase (Ambion, USA). The ligated RNA was reverse transcribed with PrimeScript (Takara), amplified with Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific, USA).