title: GSM5001850: BF1182: Ductal cell-derived tumor 2; Mus musculus; RNA-Seq
library_construction_protocol: When mice reached morbidity, pancreatic tumors were micro-dissected and flash frozen in liquid nitrogen. RNA was extracted from bulk tumors using the RNeasy midi kit (Qiagen). RNA quality was assessed using a Bioanalyzer, and only samples with an RNA integrity number (RIN) greater than 8.0 were used for library preparation. RNA-seq libraries were generated using the TruSeq RNA Sample Preparation Kit (Illumina). The mRNA was enriched using oligo(dT) beads. The mRNA is enriched using oligo(dT) beads. First, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.