RNA-Seq of Homo sapiens: PBMC CLL
Libraries are prepared using the combination of 2 kits: QIAseq FastSelect rRNA HMR Kit from QiaGen, and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina protocol according suppliers recommendations. Briefly the key stages of this protocol are successively, the removal of ribosomal RNA fraction from 80ng of total RNA where QIAseq FastSelect removal reagent is directly combined with total RNA and the library preparation-specific buffers, a fragmentation under elevated temperature to obtain approximately 300bp pieces, double strand cDNA synthesis and finally Illumina adapters ligation and cDNA library amplification by PCR for Sequencing. Libraries are sequenced on paired end 100b run on the Illumina NovaSeq. Image analysis and base calling is performed using Illumina Real Time Analysis ( 3.4.4 ) with default parameters.