GSM8256233: D3_HFD_T2; Mus musculus; RNA-Seq
RNA was isolated from mouse pancreatic neoplastic lesions at T1 (30 days; LFD mice n=3; HFD mice n=4) and T2 from orthotopic PaNIN lesion transplantation (90 days; LFD mice, HFD mice and  SEMA mice; n=4 for each experimental group) by miRNeasy micro Kit (Qiagen) combined with on-column DNase digestion (DNase-Free DNase Set, Qiagen) to avoid contamination by genomic DNA according manufacturer's instruction. Then, RNA integrity was verified by BioAnalyzer 2100 (Agilent) and only RNA samples with a RNA integrity number (RIN) > 7 were processed for cDNA library construction. Sequencing was performed on rRNA-depleted total RNA (50M PE reads, 7.5Gbp) on an Illumina HiSeq 2000 platform. The resulting raw reads were assessed for quality by FastQC and preprocessed with FASTQ Toolkit v.2.2.5 for adapter trimming and low quality (Q<30) filtering of the reads.