RNA-seq of Homo sapiens : blood serum of colorectal cancer
Sequencing libraries were prepared by NEBNext Ultra II Directional RNA Library Prep Kit for Illumina according to the manufacturers recommendations using 5 l of RNA, 18 cycles of preamplification, and NEBNext Adaptor dilution (200 ). Libraries were barcoded with NEBNext Multiplex Oligos for Illumina (all New England BioLabs, Ipswich, MA, USA). All libraries were diluted to 4 nM and pooled in equimolar ratio. Denatured and diluted PhiX Control v3 was added at 1% and RNA sequencing was performed using the NextSeq 500/550 High Output v2 Kit and the NextSeq 500 Sequencing System (Illumina, San Diego, CA, USA).