title: GSM8008863: Pancreas, KPC, mouse 48; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA (~1µg) was isolated from snap-frozen mouse pancreas tissues using TRI Reagent® Solution (Sigma) followed by on-column RNeasy® Mini Kit RNA clean-up and DNase treatment (Qiagen). Libraries were generated using an in-house multiplex RNA-seq method (MHTP, Medical Genomics Facility). Libraries were prepared using 20 ng of total RNA input. An 8 bp sample index and a 10 bp unique molecular identifier (UMI) were added during initial poly(A) priming and pooled samples were amplified using a template switching oligonucleotide. Illumina P5 was added by tagmentation by Nextera transposase and PCR and Illumina P7 was added by PCR. The library was designed so that the forward read (R1) used the standard R1 primer to sequence the cDNA in the sense direction for transcript identification. The index read (I1) used the standard i7 primer to sequence directly into the sample index and then the 10 bp UMI. Sequencing (R1 101 bp; I1 18 bp) was performed on a NextSeq 2000 (Illumina) P3 100 cycle run.