GSM5818847: 3964_5G_plus_6G_S20; Homo sapiens; RNA-Seq
Mononuclear cells were isolated from bone marrow aspirates using density centrifugation media, and stored at -80℃. After staining with CD38, CD45, CD19, and CD138, Single cell sort of MM cells was performed in 384 well plates using FACS. Libraries were prepared according to the CEL-Seq2 protocol. Briefly, mRNA from cells sorted into 384-well plates was reverse-transcribed into complementary DNA, with which 12-well samples were pooled. The pooled samples were then amplified by T7 in vitro transcription, and the amplified RNA was fragmented and used to construct final libraries by reverse transcription and PCR amplification. Every PCR product is uniquely labeled by random attachment of barcode sequences to both ends. The libraries were sequenced on Illumina HiSeq 1500 sequencers and the reads were mapped to the GRCh38 human reference genome assembly using HISAT2 (v. 2.2.1).