title: Illumina NovaSeq 6000 paired end sequencing; Selection of RNA-seq library preparation protocol for a comprehensive transcriptome analysis in T acute lymphoblastic leukemia (T-ALL)
design: Selection of RNA-seq library preparation protocol for a comprehensive transcriptome analysis in T acute lymphoblastic leukemia (T-ALL)
library_construction_protocol: T-cells (T-ALL and normal T lymphocytes) were obtained from bone marrow samples by isolation of mononuclear cells, using density gradient centrifugation, followed by immunomagnetic separation, with use of Human T Lymphocyte Enrichment Set-DM (Becton Dickinson). Total RNA was isolated using miRCURY RNA Isolation Kit Cell & Plant (Exiqon). RNA isolates were DNase treated and purified with use of RNA Clean and Concentrator Kit (Zymo Research).  Libraries were generated with Illumina TruSeq Stranded mRNA and Total RNA Library Prep Kit. Standard fragmentation conditions were used (94â€™C for 8 minutes; referred to as P1).