title: GSM5050294: CBMC_NOSTIM1; Homo sapiens; RNA-Seq
library_construction_protocol: Samples were lysed in TCL buffer (Qiagen) SmartSeq2 libraries were prepared according to the SmartSeq2 protocol described by Trombetta et al, 2014. Briefly, total RNA was purified using RNA-SPRI beads. cDNA was generated from full-length mRNA transcripts using reverse transcriptase with terminal transferase activity. Combined with a second "template switch" primer, the cDNA was constructed to have two universal priming sequences. Following preamplification, the Nextera XT library construction kit was used to prepare 96 unique indexes specific to each sample. Barcoded cDNA fragments were then pooled prior to sequencing. Sequencing was carried out using an Illumina NextSeq500 as paired- end 2x38bp to a coverage of approximately ~4 million reads per well.