title: GSM3711567: 000008512269_B09; Homo sapiens; RNA-Seq
library_construction_protocol: Naïve (CD45RA+CD28+), EM (CD45RA-CD28-) and TEMRA (CD45RA+CD28-) CD3+CD8+ were FACS-sorted (FACSAria, BD Biosciences; purity greater than 95%) from freshly isolated human PBMCs obtained from 8 healthy donors (obtained by the Establissement Français du Sang). After sorting, total RNA was extracted using the RecoverAll Total Nucleic Acid Isolation Kit (Ambion). RNA quantity was measured with a Nanodrop ND1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE) and RNA quality was assessed using an Agilent 2100 Bioanalyser. Smart-Seq2 libraries were prepared by the Broad Technology Labs and sequenced by the Broad Genomics Platform according to the SmartSeq2 protocol (Picelli et al, 2013, 2014) with some modifications (Trombetta et al, 2014). Briefly, total RNA was purified using RNA-SPRI beads, polyA mRNA was converted to cDNA and amplified cDNA was subject to transposon-based fragmentation that used dual-indexing to barcode each fragment of each converted transcript with a combination of barcodes specific to each sample. Sequencing was carried out as paired-end 2x25bp with an additional 8 cycles for each index.
attributes.tag: GEO Accession
attributes.value: GSM3711567