ncounter Analysis System For Translational Research nanostring.com FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

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1 ncounter Analysis System For Translational Research nanostring.com FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

2 Faster than qpcr. Simpler than NGS. The ncounter platform provides a simple and costeffective solution for multiplex analysis of up to 800 RNA, DNA, or protein targets from your precious samples. Save Time Expertly curated pre-formatted panels for human, mouse and non-human primate 30-minutes total hands on time without amplification, cdna or laborious library prep Sample to Publication ready figures in ~24 hours Save Sample Combine RNA, DNA, and protein panels for a comprehensive 3D Biology view of each sample Optimized performance on difficult sample types including FFPE, tissue, lysates and biofluid samples Save Resources Advanced analysis tools included with system reduce the need for Bioinformatics support Digital gene expression eliminates need for technical replicates 2

3 Molecules That Count HIGHLY MULTIPLEXED SINGLE MOLECULE COUNTING NanoString s patented molecular barcodes provide a true digital detection technology capable of highly multiplexed analysis*. CAPTURE PROBE REPORTER PROBE TARGET-PROBE COMPLEX TARGET HYBRIDIZE Two probes hybridize directly to a target molecule in solution. The Reporter Probe carries the fluorescent barcode and the Capture Probe contains a biotin moiety that immobilizes the hybridized complex for data collection. SOLUTION PHASE HYBRIDIZATION 2 PURIFY + IMMOBILIZE After hybridization, samples are transferred to an ncounter instrument which removes excess probes. Purified target-probe complexes are bound, immobilized and aligned on the imaging surface of the ncounter cartridge. EXCESS PROBES REMOVED HYBRIDIZED PROBES BIND TO CARTRIDGE COMPLEXES ARE IMMOBILIZED AND ALIGNED ON CARTRIDGE 3 COUNT Sample cartridges are scanned by an automated fluorescence microscope. Barcodes are counted for each target molecule and the data are exported as a simple CSV file. BARCODES COUNTED BARCODES COUNTS IDENTITY INSULIN 2 FOX5 3 XLSA *Single cell and DNA SNV assays require amplification prior to sample processing and data collection. 3

4 Exceptional Reproducibility and Performance Faster than qpcr, simpler than NGS Fully automated and easy-to-use, the ncounter Analysis System provides everything you need to cost-effectively complete your projects in record time. Strong analytical performance sensitive, precise and quantitative digital data Flexible samples optimized performance with most sample types including FFPE, PBMCs and FACS Single tube multiplexing up to 800 targets Quality assurance One platform for both basic research and clinical diagnostics; GMP compliant/iso 3485 certified Easy-to-use fully automated, intuitive user interface Data analysis generate publication quality figures quickly and easily with nsolver Analysis Software Amplification Free Analysis* Most ncounter assays do not require amplification of target sequence for detection and can be performed with ng of input material which is ideal for investigators working with precious samples. This amount is equivalent to a single curl of FFPE tissue and data are comparable to that generated with matched fresh-frozen material. * Single cell and DNA SNV assays require amplification prior to sample processing and data collection. 4

5 ncounter Systems ncounter SPRINT ncounter MAX ncounter FLEX AN INSTRUMENT FOR EVERY NEED integrated benchtop system for Research Labs for Core Labs and high sample throughput for Translational Labs and clinical applications ncounter LIFE SCIENCE ASSAYS ncounter ELEMENTS EXPANDABLE with Additional Prep Station No ENTERPRISE PACKAGE No PROSIGNA optional add-on No No RUNS PER DAY 2 4* 4* THROUGHPUT (LANES PER DAY) * 48-96* HANDS ON TIME 0 min 5 min 5 min * Additional Prep Station required for > 24 lanes per day. TENS OF THOUSANDS OF DATA POINTS EVERY DAY # of Genes per Run Samples per Day Data Points per Day ncounter SPRINT ( sample per lane) 800 genes 24 samples = 9,200 ncounter FLEX ( sample per lane) 800 genes 48 samples = 38,400 ncounter MAX (2 Prep Stations; sample per lane) 800 genes 96 samples = 76,800 ncounter MAX (2 Prep Stations; 4 samples per lane) 200 genes 384 samples = 76,800 INCREASE SAMPLE THROUGHPUT WITH ncounter PLEXSET CHEMISTRY # of Genes per Run Samples per Day Data Points per Day ncounter MAX (2 Prep Stations; 8 samples per lane) 24 genes 768 samples = 8,432 ncounter SPRINT (2 prep stations; 8 samples per lane) 24 genes 92 samples = 4,608 5

6 One Chemistry Many Applications NanoString s molecular barcoding technology uses color-coded molecular barcodes that can hybridize directly to many different types of target molecules. It is ideal for a range of applications requiring efficient, high-precision quantitation of hundreds of target molecules across a sample set. All ncounter assays generate high-quality results from challenging sample types, including FFPE and crude cell lysates. Gene Expression Analysis Rapidly analyze up to 800 genes simultaneously No RT, no enzyme and no amplification* Lyse and Go protocols for cells, blood and FFPE mirna Expression Analysis Multiplexed target profiling of mirna transcriptomes in a single reaction Targeted mirna discovery and validation on one platform Excellent specificity: accurately distinguish between highly similar mirnas mirge Expression Analysis Simultaneously profile mirna and mrna expression in a single reaction No RT, no amplification and fewer pipetting steps RNA mrna IncRNA Gene Fusions mirna Fusion Gene Analysis Identify fusion events without knowledge of partner genes Characterize specific fusions by probing the junction sequence Study fusions and gene expression targets in the same assay lncrna Expression Analysis High precision, digital quantification of lncrnas Analyze up to 800 lncrnas in a single reaction with no amplification Single Cell Analysis Obtain single cell sensitivity with reverse transcription and limited amplification * Low RNA Input protocol available, requires amplification 6

7 Small Sample. Big Insight. 3D BIOLOGY TECHNOLOGY Profile combinations of DNA, RNA fusion, protein and phospho-protein targets up to 800-plex from a single sample Designed for mix and match flexibility Multi-analyte pre-matched assays Minimal sample input DNA Single Nucleotide Variation (SNV) Analysis Tumor-specific panels Built-in internal controls for amplification cycle and false discovery rates (FDRs) Detects SNVs, dinucleotide variants and small InDels SNV Copy Number Variation (CNV) Analysis Custom and cancer-specific panels Internal controls including invariant genomic regions and spike-in process controls Analyzes 0 4 bi-allelic and multi-allelic CNVs CNV PROTEIN Protein and Phospho-protein Expression Analysis Multi-plex content focused on key areas in oncology research Profile 30 proteins simultaneously Customizable panels with our protein barcoding service Compatible with primary cells, fresh/frozen tissue, and FFPE 7

8 Wnt Hedgehog Apoptosis/Cell Cycle RAS PI3K STAT MAPK Notch TGF-β Chromatin Modification Transcriptional Regulation DNA Damage Control Panels to Accelerate Your Research NanoString s pre-made panels are available for a number of important pathway and research areas. All panels are created with input from industry experts and current research topics and are updated regularly. ncounter PANEL SPECIFICATIONS GENE EXPRESSION PANELS mirna TARGETED DISCOVERY PANELS DESCRIPTION NO. OF GENES DESCRIPTION NO. OF mirnas PanCancer IO Human v3 800 PanCancer Human Pathways 770 Mouse v PanCancer Human Immune Profiling 770 Rat v PanCancer Mouse Immune Profiling 770 PanCancer Human Progression 770 HUMAN KARYOTYPE PANEL Neuropathology 770 Human Myeloid Innate Immunity 770 Mouse Myeloid Innate Immunity 754 Human Immunology v2 594 DESCRIPTION Human Karyotype monitors gross chromosomal abnormalities; includes 338 individual loci to quickly monitor cells as they passage REGIONS 8 probes per arm Mouse Immunology 56 Human Kinase v2 536 CANCER CN PANEL V2 Human Inflammation v2 255 DESCRIPTION NO. OF GENES Mouse Inflammation v2 254 Human Cancer Reference 236 CNVs (~3 probes per region) commonly amplified or deleted in cancer 87 Human Stem Cell 99 Customer Assay Evaluation 47 Human Reference 8 8

9 ncounter VANTAGE 3D ASSAYS - MIX AND MATCH RNA PANELS PROTEIN PANELS DESCRIPTION NO. OF GENES DESCRIPTION NO. OF PROTEINS Adaptive Immunity 92 Innate Immunity 92 Immune Cell Profiling for cell suspensions 30 Immune Cell Signaling for cell suspensions 26 Cancer Metabolism 92 Solid Tumor for lysate 28 Intracellular Signaling 92 Solid Tumor for FFPE 26 Cellular Profiling 92 Wnt Pathways 92 Heme for lysate ** Heme for FFPE ** DNA Damage and Repair 92 MAPK-PI3K Pathways 92 Heme 92 DNA SNV PANELS DESCRIPTION NO. OF MUTATIONS Lung Fusion 63 DNA SNV Solid Tumor 04 Leukemia Fusion 42 DNA SNV Heme Inquire** ncounter VANTAGE 3D ASSAYS - PRE-MATCHED COMBINATION PANELS DESCRIPTION RNA:Protein Immune Cell Profiling for cell suspensions RNA:Protein Immune Cell Signaling for cell suspensions RNA:Protein Solid Tumor for FFPE RNA:Protein Solid Tumor for lysate DNA:RNA:Protein Solid Tumor for lysate DNA:RNA:Protein Solid Tumor for FFPE up to 800 plex DNA:Fusion:Protein Lung for lysate DNA:Fusion:Protein Lung for FFPE DNA:RNA:Protein Heme for lysate** DNA:RNA:Protein Heme for FFPE** ** Available late 207. Ask your account representative about availability. Say Hello to ndesign Gateway For a complete list of genes in each panel, visit STORE.NANOSTRING.COM 9

10 Insight from Difficult Samples ncounter assays can accept samples such as purified total RNA, raw cell or blood lysates and formalin-fixed paraffin-embedded (FFPE) extracts with no loss in precision. Even severely degraded RNA can be a viable sample input. Crude Cell Lysates 0,000 0,000 cells Three cell lysates (2,500, 5,000, and 0,000 cells) were compared to 00 ng of purified total RNA. Results using cell lysates were highly correlated with purified RNA (R2 > 0.97 for all three) and demonstrated that comparable data can be achieved with either protocol. Lysate Counts, ,000 cells 2,500 cells 0 00,000 0,000 Total RNA Counts Whole Blood Lysates 0,000 Lysate Two PAXgene -lysed whole blood replicates compared to 00 ng of matched purified total RNA. Results using blood lysates were highly correlated with purified RNA (R2 > 0.96 and R2 > 0.97) and demonstrated that high quality data can be obtained using PAXgene-lysed whole blood. (PAXgene is a trademark of QIAGEN.) Lysate Counts, Lysate ,000 0,000 Total RNA Counts Formalin-Fixed paraffin-embedded tissue 0,000 FFPE-derived and purified total RNA compared to matched purified total RNA from fresh tissue. Results using FFPE-derived tissue were highly correlated with purified RNA (R2 > 0.97) and demonstrated that high quality data can be achieved from FFPE. Lysate Counts, ,000 0,000 Total RNA Counts Expanded Options with ncounter Low RNA Input Kit (-0 ng) The ncounter Low RNA Input Kit enables high quality gene expression profiling of up to 800 gene targets from as little as ng of sample. The kit is optimized for use with RNA from Formalin Fixed Paraffin Embedded (FFPE) tissue as well as crude cell lysates. Additionally, the kit can be utilized in the study of low expressing genes. The streamlined, user friendly workflow and reliable results enable gene expression studies of small samples or low expressing genes to be completed quickly and efficiently. Fold Change Low RNA Input ( ng) UHR/HBR Fold Change Standard assay (00 ng) UHR/HRB 3 5 * Sample inputs of ng can be achieved with the ncounter Low RNA Input kit. 0

11 OR3443 OR3457 OR3458 OR3452 OR024 OR298 OR04 OR323 OR3242 OR3567 OR3005 OR304 OR2969 OR2968 OR3550 OR288 OR007 OR004 OR07 OR020 OR003 OR03 OR00 OR006 OR002 OR3465 OR3420 OR3379 OR3466 OR3392 OR3398 OR2892 OR3220 OR3247 OR2899 OR3542 OR2975 OR02 OR3249 OR2858 OR383 OR297 OR3273 OR2890 OR2862 OR3208 OR2868 OR03 OR3243 OR324 OR2976 OR2878 OR384 OR2686 OR2700 OR3397 OR3470 OR2707 OR2705 OR272 OR3004 OR3449 OR3468 OR2908 OR3236 OR3380 OR3435 OR3440 OR347 OR2952 OR2955 OR2888 OR3437 OR07 OR2925 OR297 OR296 OR2963 OR2965 OR2854 OR3422 OR3462 OR2973 OR29 OR3387 OR3476 OR2859 OR2967 OR3478 OR2872 OR2856 OR3229 OR05 OR342 OR06 OR344 OR2893 OR3228 OR344 OR3403 OR2897 OR2895 OR2853 OR3258 OR3022 OR272 OR3383 OR338 OR3453 OR2903 OR290 OR3489 OR3425 OR3444 OR34 OR3385 OR286 OR3275 OR08 OR3433 OR3408 OR3442 OR2905 OR399 OR2962 OR3233 OR3269 OR2879 OR3226 OR2974 OR3268 OR2870 OR386 OR2904 OR2877 OR387 OR2693 OR306 OR3456 OR3430 OR02 OR09 OR3464 OR3009 OR2682 OR322 OR2923 OR292 OR390 OR3544 OR3244 OR275 OR3076 OR2866 OR2958 OR325 OR2865 OR023 OR2863 OR294 OR3209 OR2966 OR3224 OR2884 OR397 OR3245 OR2959 OR2696 OR303 OR3206 OR295 OR2889 OR3276 OR3399 OR3434 OR2902 OR2864 OR3054 OR3025 OR2926 OR3230 OR3402 OR2857 OR2694 OR3026 OR3203 OR2894 OR3223 OR2907 OR2852 OR3545 OR3390 OR3482 OR32 OR2876 OR2887 OR329 OR3388 OR3473 OR296 OR3506 OR2855 OR3222 OR3432 OR00 OR388 OR287 OR2954 OR2957 OR346 OR3393 OR3474 OR3436 OR2882 OR2873 Powerful Data Analysis VISUALIZE RESULTS WITH nsolver ANALYSIS SOFTWARE nsolver Analysis Software is an integrated analysis platform for storage, custom QC, and custom normalization of ncounter data. Generate highly-customized exports, basic statistical outputs, and publication-quality figures quickly and easily with the no incremental cost. Recommended quality control on samples/lanes Tunable normalization and fold-change measurements Statistical significance testing Compatible with standard analysis programs including; Ingenuity Pathway Analysis, Partek Genomics Suite, BioDiscovery Nexus Copy Number, Advaita ipathwayguide Subtype Binding.Density Scanned.Date Scanned.Date Binding.Density Subtype OR3443 OR3457 OR3458 OR3452 OR024 OR298 OR04 OR323 OR3242 OR3567 OR3005 OR304 OR2969 OR2968 OR3550 OR288 OR007 OR004 OR07 OR020 OR003 OR03 OR00 OR006 OR002 OR3465 OR3420 OR3379 OR3466 OR3392 OR3398 OR2892 OR3220 OR3247 OR2899 OR3542 OR2975 OR02 OR3249 OR2858 OR383 OR297 OR3273 OR2890 OR2862 OR3208 OR2868 OR03 OR3243 OR324 OR2976 OR2878 OR384 OR2686 OR2700 OR3397 OR3470 OR2707 OR2705 OR272 OR3004 OR3449 OR3468 OR2908 OR3236 OR3380 OR3435 OR3440 OR347 OR2952 OR2955 OR2888 OR3437 OR07 OR2925 OR297 OR296 OR2963 OR2965 OR2854 OR3422 OR3462 OR2973 OR29 OR3387 OR3476 OR2859 OR2967 OR3478 OR2872 OR2856 OR3229 OR05 OR342 OR06 OR344 OR2893 OR3228 OR344 OR3403 OR2897 OR2895 OR2853 OR3258 OR3022 OR272 OR3383 OR338 OR3453 OR2903 OR290 OR3489 OR3425 OR3444 OR34 OR3385 OR286 OR3275 OR08 OR3433 OR3408 OR3442 OR2905 OR399 OR2962 OR3233 OR3269 OR2879 OR3226 OR2974 OR3268 OR2870 OR386 OR2904 OR2877 OR387 OR2693 OR306 OR3456 OR3430 OR02 OR09 OR3464 OR3009 OR2682 OR322 OR2923 OR292 OR390 OR3544 OR3244 OR275 OR3076 OR2866 OR2958 OR325 OR2865 OR023 OR2863 OR294 OR3209 OR2966 OR3224 OR2884 OR397 OR3245 OR2959 OR2696 OR303 OR3206 OR295 OR2889 OR3276 OR3399 OR3434 OR2902 OR2864 OR3054 OR3025 OR2926 OR3230 OR3402 OR2857 OR2694 OR3026 OR3203 OR2894 OR3223 OR2907 OR2852 OR3545 OR3390 OR3482 OR32 OR2876 OR2887 OR329 OR3388 OR3473 OR296 OR3506 OR2855 OR3222 OR3432 OR00 OR388 OR287 OR2954 OR2957 OR346 OR3393 OR3474 OR3436 OR2882 OR2873 SubtypeD SIMPLE, ADVANCED DATA ANALYSIS ncounter Advanced Analysis is a free, wizard-based addon to nsolver Analysis Software for deeper data insights based on robust R statistics. Examine experimental trends, identify pathway-specific responses, and profile immune cell populations in shareable HTML reports. Support for all mrna and protein CodeSets, including custom reagents and panels Quick Analysis option for one-click data QC, normalization, and differential expression testing Automatic incorporation of biological annotations and logical defaults for each panel log0(p value) KIT CCNA2 SPRY2 PKMYT UBE2T LIFR H2AFX MAML2 ERBB2 NTRK2 ITGA7 CCNB IGF HISTH3H ILRA EZH2 COMP TTK INHBA PDGFD PPARG E2F HISTH3B RUNXT LEPR PTTG2 CDKNC JUN FANCA CDKN2C RAD5 CDC25C CDC6 ID4 CCNE2 XPA COLA MYD88 CDC25A p < 0.0 p < 0.00 SFRP Log(fold change) Data analysis services for large projects are available. For more information please contact: DAS@nanostring.com

12 Custom Customize Codeset your Design Solution DESIGN A CUSTOM CODESET SPECIFIC TO YOUR RESEARCH Standard chemistry enables processing of up to 96 samples/day x 800 (depending on system) PlexSet chemistry enables sample multiplexing of up to 8 samples per lane, increasing sample throughput CUSTOMIZE A PANEL Add up to 30 additional genes or a collection of specific controls to make your panel unique to that experiment. SELECT 2 PROBE 3 CUSTOMER 4 GENES DESIGN REVIEW Submit your RefSeq IDs for up to 800 target genes to NanoString. LEAD TIME Customer-defined NanoString designs probes then creates and sends a Design Report. LEAD TIME Custom GEx: 3-5 days Custom CNV: 0-5 days Customer reviews and approves Design Report. LEAD TIME Customer-defined MANUFACTURE AND SHIP NanoString manufactures and ships CodeSet to customer. LEAD TIME 3-5 weeks (dependent on gene number and scale) Say Hello to ndesign Gateway For a complete list of genes in each panel, visit STORE.NANOSTRING.COM 2

13 One Instrument from Lab to Clinic CLINICAL DIAGNOSTICS CAPABILITIES Our experience developing, testing and marketing Prosigna demonstrates our commitment to establishing ncounter as a truly multi-purpose platform for research and clinical use. MANUFACTURING Reagent production Process validation GMP and ISO 3485 certification DEVELOPMENT CLINICAL Assay development Algorithm training Software development Assay SOP transfer Medical affairs Site monitoring COMMERCIAL REGULATORY Reimbursement planning Marketing Sales Pre-IDE IDE submission FDA 50(k) clearance The ncounter FLEX is manufactured under GMP guidelines and ISO 3485 to ensure quality and compliance with international standards. In Life Sciences mode, ncounter FLEX can perform all ncounter Life Science and ncounter Elements assays for research use. Diagnostics mode provides a secondary interface for running diagnostic assays such as the Prosigna Breast Cancer Prognostic Gene Signature Assay. VALIDATE YOUR OWN IVD ASSAYS ncounter Elements are a set of reagents and consumables that are registered with the FDA and are intended for use with ncounter technology to enable the end-user to validate diagnostic assays. REPORTER TAG Selected probes procured by end user UNIVERSAL CAPTURE TAG A unique tag sequence PROBE A** PROBE B** is assigned to each target sequence 5 3 NUCLEIC ACID TARGET 3

14 Your Trusted Advisor A team of highly dedicated support and service personnel are available to ensure your success with all ncounter products. Field Applications Scientists and Technical Services Scientists are highly trained experts who are available to assist you before, during and after your project. Field Service Engineers will ensure your system is operating at peak performance and will qualify your instrument as needed through a variety of service offerings. LET US HELP YOU MAXIMIZE THE BIOLOGICAL INFORMATION CONTAINED WITHIN YOUR SAMPLES THROUGH THESE PRODUCTS AND SERVICES: CONSULTATION TRAINING CUSTOM SEMINARS Experimental Design Strategies New System Training Technology Overviews Application Design New Application Training New Technologies Product Selection Strategies Data Analysis/Advanced Analysis Training TROUBLESHOOTING SERVICE DATA ANALYSIS Assay Optimization Tiered Service Contracts nsolver and Basic Analysis Support Log File Interpretation IQ/OQ/PQs Data Analysis Project Services (for fee) Many thanks for taking care of us and your support. Again, I have to say that the NanoString Support is superb, and other companies should use it as a role model. NanoString customer Contact Support support@nanostring.com 4

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16 For more information, please visit nanostring.com NanoString Technologies, Inc. 530 Fairview Avenue North Seattle, Washington 9809 T (888) F (206) nanostring.com info@nanostring.com Sales Contacts United States us.sales@nanostring.com EMEA: europe.sales@nanostring.com Asia Pacific & Japan apac.sales@nanostring.com Other Regions info@nanostring.com FOR RESEARCH USE ONLY. Not for use in diagnostic procedures. NanoString, NanoString Technologies, the NanoString logo, mirge, Molecules that Count, ncounter, ndesign, nsolver, ncounter Elements, IO 360, 3D Biology, and Vantage 3D are trademarks or registered trademarks of NanoString Technologies, Inc., in the United States and/or other countries. Any other trademark that appears in this document is the property of its respective owner. USNS_PM093 AUG 207