3' adapters were ligated and 5' ends were modified, followed by 5' adapter ligation. The biotin-labelled RNA was extracted using Trizol, hydrolyzed using NaOH and enriched using streptavidin beads. The nascent RNA generated using biotin-run-on reaction of nuclei was used for library preparation. The cells were permeabilized to prepare nuclei. After 6 days of selection in 1 mg/mL Geneticin, the stably integrated cells, both control and experimental, were treated with 1 ug/mL Doxycycline to induce the expression of miRNAs for seven days.įlp-In™ T-REx™ 293 (HEK293) cells were cultured in DMEM containing 10% FBS, 1% Pen/Strap and 0.1% Zeocin. The cells were transduced with lentiviral vectors with or without miRNA expression cassette.
GEO help: Mouse over screen elements for information.Įmpty vector control rep 2 (batch 1)
