Exiqon micro RNA array services Niels M Frandsen

Exiqon micro. RNA array services Niels M Frandsen Product Manager

Agenda • micro. RNA array services • • Introduction to array services Workflow Follow up sales Key selling points Competitor analysis Battle cards – how to sell against the competition Things to come

micro. RNA profiling services • • Established spring 2006 4 scientists and 4 lab technicians Service includes: • RNA QC • Hybridization, scanning and image analysis • Data analysis: Normalisation, heatmap, PCA plots, statistics etc Lab: • Hybridization stations: Tecan HS 4800 Pro • Array scanner: Agilent G 2505 B Microarray Scanner System • Scanning takes place in ozone free lab to prevent bleaching flourescent dyes • ~ 500 customers (including Big Pharma) from all over the world • ~ 6000 arrays processed • ISO 9001: 2000 certified • October 2008: q-RT-PCR in Services • Roche Light. Cycler 480 (LC 480) • Eppendorf ep. Motion 5075 LH

Rapid growth of arrays service business The number of slides processed doubled in 2008 compared to 2007. #slides processed 3257 in Services 1632 2007 2008 Most significant growth comes from university hospitals – clinical samples >30% of all samples analyzed. This group also features the largest number of samples per project!

TRAINING Profile of our customers Potential customers for services are: 1) Clinical research and all stages of drug development. Well funded! 2) Academic labs that need some initial results fast in order to find out if they want to work with mi. RNA profiles. 3) Academic labs that have experience with micorarrays and understand the advantages in terms of saved time and reliability that a professional service offers. 4) Often limited funding: Service experiments are expensive! Academic labs that have no experience with microarray analysis and/or that do not want to set everything up (buy or borrow equipment, learn and optimize protocols etc. ) Hospitals – doctors investigating potential correlation between micro. RNA profiles in patients and f. ex response to clinical treatment regime – cancer. 5) Biotech and pharmaceutical companies that have the funds to outsource experiments performed using specialized techniques, and who want reliable results fast. Possibilities: - initial experiments to see if mi. RNA may be interesting in their system - large scale screening experiments for biomarker or target identification (esp. related to cancer and other diseases).

TRAINING mi. RNA micro array services workflow Consultation Sample submission RNA QC Labeling and Hybridization Data collection Normalisation Basic analysis Final report

Workflow - Consultation Topics discussed: • Type of experiment – what are the questions? - which approach is best suited? common reference, direct comparison, single (color/dual color analysis) – pilot experiments needed? • Type of samples – cell culture, tissue, FFPE, blood, enriched etc. • We recommend total RNA – our platform is not affected by m. RNA • With enriched RNA: • we get a skewed profile – each manipulation introduces a bias • we cannot perform QC • we cannot quantitate on the Nanodrop. • we request 100 ng corresponding total RNA for QC and quantitation purposes. • Amount of sample available/needed • Number of biological replicates • Sample submission form

Workflow – Consultation Amount of RNA needed (complex issue): • What we use the RNA for: • 100 ng for QC (optional) • Xng for Hy 3 • Xng for common reference (optional – can be replaced with alternative reference) • 2 x. Xng backup for rerun (optional) • General recommendations: • For most practical purposes 2002000 ng is enough • Sample volumes: 10 -30µl • Sample concentration: preferably >400 ng/ul to avoid speed vac. Not below 50 ng/µl! Low input RNA comes at the expense of reduced call rate

Workflow – Consultation Biological replicates • • • The number of biological replicates needed depends on the sample type and how different the groups the are and how reproducible the RNA sampling and extraction is performed. In general the variation is greater in tissue samples compared to cell culture samples. General recommendations: • Cell cultures: minimum 4 replicates, preferably 6 • Tissue samples: minimum 6 replicates • Pooling is only recommended for pool sizes >4 and number of pools >3

Workflow – Consultation Sample submission form Customer sends filled in SSF to: services@exiqon. com The technical application specialist sends approval by e-mail to customer Cc. distributor An order confirmation is sent from Exiqon to distributor Customer prints approved SSF and sends a signed copy along with the samples. NB: Samples may only be sent to Exiqon upon approval of SSF!!

Workflow – Consultation Sample submission form

Workflow – Consultation Sample submission form

Workflow – Consultation Sample submission form

Workflow – Consultation Sample submission form

Workflow: Submission of samples • RNA samples should be shipped on dry ice - at least 3, 5 kg is recommended for a transit time of 2 -3 days. • Please ensure that your samples are labeled clearly (as listed on the sample submission form) using a permanent marker. • Ship with a signed copy of the sample submission form • Samples should be shipped using an international courier service such as Fed. Ex. • If you are shipping from outside Europe, please only send your samples out Monday or Tuesday to avoid weekend deliveries. • Please ship samples to: Christian Glue Services Exiqon A/S Bygstubben 10 DK-2950 Vedbaek Denmark Phone +45 4565 0432

Workflow: RNA quality control Recommendations: RIN values > 7 (Bioanalyzer) OD 260/280 > 1. 7 (Nanodrop) OD 260/230 > 1. 7 (Trizol purication) (Nanodrop) RNA QC report sent to customer by e-mail Cc. distributor

Workflow: Labeling, hybridization, scanning and image analysis Customer and distributor are notified by e-mail if delays or technical problems occurs during these steps

Workflow: Final report Upon completion of final report the customer receives an e-mail with a link to a Zip file on our secure Webserver. The distributor is notified by e-mail that final report has been delivered. The Zip file contains all documents relevant to the project: RNA QC, raw data files, summary report and extensive Excel file with normalized data from each slide as well as the various types of advanced data analysis. Upon delivery of final report the customer can communicate directly with the responsible array scientist. Additional data analysis <1 hr work is included in the price.

Follow up sales: microarray experiments lay the foundation for years of downstream work! microarray services PCR validation - services Systematic follow up on microarray service customers 2 – 4 weeks after final report Detection probes - ISH Functional analysis – KD probes In vivo KD

Key selling points • • Exiqon mi. RCURY LNA™ micro. RNA microarrays • Superior sensitivity/specificity • Flexible for input amounts of total RNA 200 ng – 2000 ng • Works with FFPE – robust towards m. RNA a r. RNA degradation products • High coverage – mi. RBase 11 + 435 mi. RPlus • We offer single and dual color analysis • Robust to 3’ heterogeneity • Platform on GEO and Array Express facilitating publication of results State of the art facilities – automated hybridization stations, advanced scanners operated in ozone free environment to prevent bleaching of fluorescent dies Highly skilled and experienced array scientists Initial consultation with array expert that can offer help with experimental design Direct contact with the array scientists performing the experiments Comprehensive report and truly customized data analysis including clustering, heatmap, PCA, Annova, T-test etc Services scientists are in daily contact with our R&D and QC array scientists – intimate knowledge of the array platform We use large batches of reagents and when possible don’t perform projects on different array batches to minimize technical variation 20

Thank you for your attention 21 17 March 2018 21