Mutant TP53 in Duodenal Samples of Pancreatic Juice From Patients With Pancreatic Cancer or High-Grade Dysplasia  Mitsuro Kanda, Yoshihiko Sadakari, Michael Borges, Mark Topazian, James Farrell, Sapna Syngal, Jeffrey Lee, Ihab Kamel, Anne Marie Lennon, Spencer Knight, Sho Fujiwara, Ralph H. Hruban, Marcia Irene Canto, Michael Goggins  Clinical Gastroenterology and Hepatology  Volume 11, Issue 6, Pages 719-730.e5 (June 2013) DOI: 10.1016/j.cgh.2012.11.016 Copyright © 2013 AGA Institute Terms and Conditions

Figure 1 (A) Prevalence of TP53 mutation by diagnostic group. (B and C) Shifted melt curves, difference curves, and Sanger sequencing of digital HRM–positive PCRs from juice samples from subjects with pancreatic cancer (arrows indicate mutations). (B) Case 40, and (C) case 29 (Table 2). Clinical Gastroenterology and Hepatology 2013 11, 719-730.e5DOI: (10.1016/j.cgh.2012.11.016) Copyright © 2013 AGA Institute Terms and Conditions

Figure 2 (A) Mutant TP53 juice concentrations (mean mutation score) (y-axis) from subjects with pancreatic cancer and high-grade precursor lesions (x-axis) (*P = .0191). (B) TP53 mutation detected in a pancreatic juice sample from a subject with PanIN-3 (case 4, Table 3). Shifted melt curves, difference curves, and sequencing of digital HRM PCRs (arrows indicate mutations). Clinical Gastroenterology and Hepatology 2013 11, 719-730.e5DOI: (10.1016/j.cgh.2012.11.016) Copyright © 2013 AGA Institute Terms and Conditions

Supplementary Figure 1 Representative examples of shifted melting curves, difference curves, and Sanger sequencing. Identical TP53 mutations detected in both primary cancer tissue and juice samples by digital HRM and sequencing samples. (A) Exon 5; homozygous substitution, (B) exon 6; heterozygous substitution, (C) exon 7; deletion. Arrows indicate detected TP53 mutations. WT, wild-type control. Clinical Gastroenterology and Hepatology 2013 11, 719-730.e5DOI: (10.1016/j.cgh.2012.11.016) Copyright © 2013 AGA Institute Terms and Conditions

Supplementary Figure 2 Results of stepwise dilution test in (A) exon 7 and (B) exon 8. Shifted melting curves and difference curves with distinct differences from wild-type controls were shown in all 0.1% to 10% dilution levels. (C) Scatter-plot graph of actual and estimated number of positive wells. The R2 value was 0.985, indicating high accuracy of the HRM and dilution series. Clinical Gastroenterology and Hepatology 2013 11, 719-730.e5DOI: (10.1016/j.cgh.2012.11.016) Copyright © 2013 AGA Institute Terms and Conditions