GSM5088557: CRC_N_LI_M_93; Homo sapiens; RNA-Seq
surgical resection tissue was stored in RNAlater (Qiagen) upon collection until further processing. Tissue was homogenised using the soft tissue homogenizing CK14 kit (Precellys, Stretton Scientific 03961) in 300µl of RLT lysis buffer (Qiagen) and 20µM DTT (Sigma). RNA was isolated using the Qiagen Mini kit with a DNA digestion step (Qiagen). Sequencing libraries were prepared using either the QuantSeq 3' mRNA-Seq FWD Library Prep Kit (Lexogen) for whole tissue samples or the Smart-seq2 protocol (Picelli et al., 2013) for bulk and cultured cell populations (with in-house indexing primers) RNA-sequencing, Libraries were sequenced using an Illumina HiSeq4000 with 75bp paired-end sequencing (Lamble et al., 2013).