GSM8403574: iPSC-CLC_NoRFP-SampleC 2; Homo sapiens; RNA-Seq
total RNA was extracted using TRIZOL LS by Single Cell Discoveries B.V. The sorted cells were lysed in TRIzol LS (Invitrogen, #1029602) and stored at -80℃. After RNA extraction, bulk RNA sequencing was performed using a modified version of the CEL-Seq protocol(Hashimshony et al. 2012) by Single Cell Discoveries B.V. (Utrecht, The Netherlands).  All extracted and purified RNA samples were assessed by Agilent Tapestation and passed the quality control with high quantity and quality (with RNA integrity number (RIN) scores > 6). Subsequently, these mRNA samples were barcoded with CEL-seq primers during reverse transcription after which they were pooled for second strand synthesis. Resulting cDNA were pooled into one library and amplified using in vitro transcription reaction (IVT). The amplified RNA sequencing libraries were prepared with Illumina TruSeq small RNA primers (Illumina), for paired-end sequencing on the Illumina Novaseq™ X Plus, with a 10B 100 cycles Illumina kit (read 1: 26 cycles, index read: 6 cycles, read 2: 60 cycles). Read 1 was used to identify the Illumina library index and CEL-seq2 sample barcode.