title: GSM8512806: Akata Induced, 4; Homo sapiens; human gammaherpesvirus 4; RNA-Seq
library_name: GSM8512806
library_construction_protocol: RNA extraction Cells were resuspended in 1ml of TRIzol Reagent (Invitrogen, cat no. 15596026) and processed according to the manufacturer's protocol. RNA pellets were resuspended in 42 μl of ddH2O. 5 μl of 10× DNase buffer and 3 μl of DNase I (New England Biotechnology, cat no. M0303L) were added, and tubes were incubated at 37°C for 15 min. DNase I was removed using the Monarch RNA Cleanup Kit (New England Biotechnology, cat no. T2030L). 1μg of RNA was reverse transcribed using LunaScript (New England Biotechnology, cat no. M3010), according to the manufacturer's protocol. RNA quality was assessed using the Agilent 2100 Bioanalyzer (Agilent, Serial No. DE54107860) with the Agilent RNA 6000 Nano Kit (Agilent, Cat. No. 5067-1511) to ensure RNA integrity numbers of greater than 9.5. Oxford nanopore long read sequencing ONT (Oxford Nanopore Technologies) sequencing was performed using the PromethION platform (Oxford Nanopore Technologies plc.) at the Biotechnology Center, University of Wisconsin-Madison. The sequencing run was conducted on a FLO-PRO002 flow cell with the SQK-PCB109 barcoding kit according to the manufacturer's instructions. The experiment was run for a total duration of 72 hours.