GSM5221055: Fibro-iPSC.2_IFN; Homo sapiens; RNA-Seq
Total RNA extraction was done using TRIzol (Invitrogen) and further purification with the RNeasy Micro Kit (QIAGEN) from 3 x 10^6 Fibro-iPSC.2 and Fibro-iPSC.2_IFN cells, and from 5 x 10^6 cells for all other samples. cDNA library preparation was done using 1000 ng RNA for hiPSC22_IFN and 4000 ng RNA for all other samples, using the KAPA Hyperprep RNAseq stranded kit (KAPA) with polyA capture. 9 and 7 PCR cycles for hiPSC22_IFN and all other iPSC samples, respectively, were used for library amplification. All libraries were diluted to 10 nM and normalized by qPCR using the KAPA library quantification kit (KAPA). Libraries were pooled to equimolar concentration. Sequencing was performed with the Illumina Nextseq500 using the Nextseq High Output 150 cycles (2x80bp for hiPSC22 and hiPSC22_IFN, and 2x75bp for all other iPSCs) using 2 pM of the pooled libraries.