title: RNA-seq of pancreatic cell lines AsPC1 and BxPC3 treated with Tumor Treating Fields (TTFields) compared to control samples
library_construction_protocol: Samples were fresh frozen in TRIzol Grown in a 37 °C humidified incubator in RPMI1640 media supplemented with 10% FBS. All media were further supplemented with 2 mM L-glutamine and 50 μg/ml penicillin/streptomycin. TTFields were applied to cells using the inovitroTM system as previously described (Kirson, E.D., et al., Disruption of cancer cell replication by alternating electric fields. Cancer Res, 2004. 64(9): p. 3288-95.) With frequency of 150 kHz and intensity of 1.7 V/cm RMS for 24 and 48 hours.  RNA was purified with TRI reagent as previously described [ Rio, D.C., et al., Purification of RNA Using TRIzol (TRI Reagent). Cold Spring Harbor Protocols, 2010. 2010(6): p. pdb.prot5439. ]   For cDNA libraries preparation, the polyA fraction (mRNA) was purified, followed by fragmentation and generation of double-stranded cDNA. Agencourt AMPure XP beads (Beckman Coulter) were used for cleanup, followed by end repair, UMIs addition, adapter ligation and PCR amplification steps.