Pre-validated Sequencing Assays Sequence assays were developed and validated in collaboration with leading researchers at the Dana Farber Cancer Institute. Customers supply genomic DNA and Agencourt performs nested PCR and bi-directional sequencing followed by automated PolyPhred analysis. Flexible Analysis Packages Agencourt has optimized the latest version of PolyPhred to produce highly accurate SNP identification with low false positive rates and low false negative rates. We have developed a proprietary algorithm to deploy different PolyPhred stringencies for specific sequence contexts in any given amplicon. This dynamic PolyPhred deployment has increased the PolyPhred accuracy to above 90%. PolyPhred analysis, even with optimization, has limitations identifying insertions and deletions. Agencourt offers manual analysis of these automated reports to eliminate any remaining false positives and to identify insertions and deletions. Basic manual analysis includes identification of insertion and deletion start sites, while advanced manual analysis also includes annotation of all inserted and deleted sequences within each sample. Agencourt's proprietary SNP Scoring Software allows rapid identification of: Intronic and exonic SNPs Amino acid changing SNPs dbSNP novelty of SNPs Patient genotypes

SNPs, insertions and deletions in over 92 genes including EGFR are accurately detected using our pre-validated sequencing assays and flexible analysis packages. Optional manual analysis eliminates false positives and identifies and annotates insertions and deletions as shown in exon 19 of EGFR shown above. Automated software annotates both previously known (dbSNP) and newly discovered SNPs. Intron and exon boundries are annotated in addition to any amino acid changing SNPs and the respective amino acid change.

The Dana-Farber Cancer Institute has licensed the sequencing assays listed below to Agencourt*. Please contact your local sales representative for exon coverage of the genes. If your gene of interest is not listed below, we feature a full SNP Discovery program for polymorphism assay development and specimen screening.

ABL1 ABL2(ARG) ACK1 AKT1 AKT2 AKT3 ALK AXL BLK BMX BRAF BTK C20orf64 CSF1R(FMS) CSK EGFR ERBB2(HER2) ERBB3(HER3) ERBB4(HER4)

EPHA1 EPHA2 EPHA3 EPHA4 EPHA7 EPHA8 EPHB1 EPHB2 EPHB3 EPHB4 EPHB6 FER FES FGFR1 FGFR2 FGFR3 FGFR4 FGR FLT1

FLT3 FLT4 FRK FYN HCK IGF1R INSR ITK(LYK) JAK1 JAK2 JAK3 KDR KIT LCK LMR LTK LYN MATK(CTK) MERTK(MER)

MET(HGFR) MST1R(RON) MUSK NTRK1(TRKA) NTRK2(TRKB) NTRK3(TRKC) PDGFRA PDGFRB PDPK1 PIK3CA PTEN PTK2(FAK1) PTK2B(FAK2) PTK6(BRK) PTK7(CCK4) PTK9 PTK9L RET ROR1

ROR2 ROS1 RYK SRC SYK TEC TEK(TIE2) TIE TNK1 TP53 TXK TYK2 TYR03 YES1 C20orf64 DKFZp761P1010

* The Dana-Farber Cancer Institute has licensed the Biological Materials for all genes except EGFR to Agencourt for the purpose of providing licensed services.