Background: Pancreatic adenocarcinoma (PA) has one of the worst prognoses of all malignancies prompting the search for oncogenic mutations that might be targets for therapy. Methods: We conducted a retrospective analysis of patients with PA who had next-generation sequencing (NGS). Key genes included were APC, ATM, BRAF, BRCA1, BRCA2, FLT3, NOTCH1, MET, FGFR4, MAPK1, MAP2K2, IDH1, IDH2, RET, JAK2, KRAS, NRAS, PTEN, EGFR, PIK3CA, PDGFRA, RUNX1, KIT, ALK, and TP53. Fluorescence in-situ hybridization (FISH) for EGFR gene amplification and MLL and ALK gene rearrangement was performed in a subset of patients. Variants were annotated with data incorporated from dbSNP, ClinVar and COSMIC. In-silico predictions were performed where applicable using PROVEAN (Protein Variation Effect Analyzer) and SIFT, and/or Polyphen (Polymorphism Phenotyping v2). Results: Among 45 patients with NGS, 31 patients (68.9%) had KRAS mutations and 14 patients (31.1%) were wild-type for KRAS. Of 29 patients whose tumors underwent FISH, 3 patients had EGFR gene amplification, 1 patient had both EGFR gene amplification and ALK gene rearrangement, and 1 patient was positive for the MLL gene rearrangement. There were novel mutations not previously described in the COSMIC database, and at least 8 were predicted to be deleterious or damaging. There was a novel missense variant observed in BRCA1 and several missense variants of unknown significance in BRCA2. A novel variant was found in RET, predicted by Polyphen as probably damaging and by Provean as neutral. A novel variant in PIK3CA was predicted to be deleterious by Provean/SIFT. Possible targetable alterations include mutations in NOTCH1, JAK2, ATM, FLT3, PTEN and EGFR, among others Of the 31 patients with mutant KRAS, 21 had resectable PA with known follow-up, with a median overall survival of 21.5 months (8.4-58.9). In the wild-type KRAS patients, 9 of the 14 patients had resectable PA with known follow-up, with a median overall survival of 17.3 months (6.2-91.6). Conclusions: NGS of PA confirmed common mutations and also revealed genomic alterations that may be potentially responsive to available targeted agents. The discovery of novel mutations PA warrants further investigation.