title: GSM4822349: HEK293T non-infecetd Rep2; Homo sapiens; RNA-Seq
library_construction_protocol: At 12, 18 and 24hr post-infection, the cells were lysed in Trizol (Thermo, 15596026) ,and the total RNA was isolated using standard phenol chloroform extraction. Standard Illumina TruSeq Stranded mRNA (LT) was performed using 500 ng of total RNA (illumina, FC-122-2101). Oligo-dT beads were used to capture polyA-tailed RNA, followed by fragmentation and priming of the captured RNA (8 minutes at 94ºC). Immediately first strand cDNA synthesis was performed. Second strand cDNA synthesis was performed using second strand marking master and DNA polymerase 1 and RNase H. cDNA was adenylated at the 3' ends followed immediately by RNA end ligation single-index adapters (AR001-AR012). Library amplification was performed for 12-15 cycles under standard illumina library PCR conditions. Library quantitation was performed using Agilent 2200 TapeStation D1000 ScreenTape (Agilent, 5067-5582). RNA sequencing was performed on the NextSeq 550 System using a NextSeq V2.5 High Output 75 cycle kit (illumina, 20024906) or 150 cycles kit (illumina, 20024907) for paired-end sequencing (70nt of each end). PolyA enrichment followed by Illumina TrueSeq adaptor ligation