miRNA and mRNA sequences in pancreatic cancer

Extracellular vesicle (EV) RNA was isolated from PDAC patient exosomes using TRIzol LS, followed by chloroform phase separation and GlycoBlue-assisted purification. RNA quality was assessed on an Agilent Bioanalyzer 2100 using the Small RNA and RNA Pico kits. Small RNA libraries were prepared with the NEBNext Small RNA Library Prep kit (12-15 PCR cycles) and size-selected at 140-160 bp using BluePippin. For mRNA, libraries were prepared using the SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian with rRNA depletion (12 PCR cycles). Pooled libraries were sequenced on an Illumina NovaSeq 6000 S4 platform (PE150 for mRNA; single-end for small RNA).