NextSeq 500 paired end sequencing; Heterogeneous effects of calorie content and nutritional components underlie dietary influence over pancreatic cancer susceptibility in mice and humans
Heterogeneous effects of calorie content and nutritional components underlie dietary influence over pancreatic cancer susceptibility in mice and humans
RNAseq was performed on tumors extracted from TAg+ mice fed control, low sugar, high protein, 10% fat, 30% fat, high glucose, or sugar-free diets. Mice were assessed for tumour formation by palpation and dissected tumours were selected for similar size before sample freezing. Total RNA was isolated using RNeasy Mini kit (Qiagen). RNA concentration and purity were determined spectrophotometrically using the Nanodrop ND-1000 (Nanodrop Technologies) and RNA integrity was assessed using a Bioanalyser 2100 (Agilent).  3′mRNA-seq library preparation and transcriptome analysis was performed using the QuantSeq 3′mRNA-Seq Library Prep Kit for Illumina and QuantSeq data analysis workflow.