exiqon microrna array services niels m frandsen product manager
TRANSCRIPT
Exiqon microRNA array services
Niels M FrandsenProduct Manager
Agenda
• microRNA 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
microRNA 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 HS4800 Pro
• Array scanner: Agilent G2505B 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 LightCycler 480 (LC 480)
• Eppendorf epMotion 5075 LH
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Rapid growth of arrays service business
The number of slides processed doubledin 2008 compared to 2007.
Most significant growth comes from universityhospitals – clinical samples >30% of all samples analyzed. This group also features the largest number of samples per project!
Potential customers for services are:
1) 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.)
2) Academic labs that need some initial results fast in order to find out if they want to work with miRNA 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) Hospitals – doctors investigating potential correlation between microRNA 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 miRNA may be interesting in their system- large scale screening experiments for biomarker or target identification (esp. related to cancer and other diseases).
Often limited funding: Service experiments are expensive!
TRAINING
Clinical research and all stages of drug development.Well funded!
Profile of our customers
miRNA micro array services workflow
ConsultationConsultation
RNA QC
Labeling and Hybridization
Data collectionNormalisationBasic analysis
Final report
Sample submission
TRAINING
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 mRNA
• 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 100ng corresponding total RNA for QC and quantitation purposes.
• Amount of sample available/needed
• Number of biological replicates
• Sample submission form
Workflow – ConsultationAmount of RNA needed (complex issue):
• What we use the RNA for:
• 100ng for QC (optional)
• Xng for Hy3
• Xng for common reference
(optional – can be replaced with alternative reference)
• 2xXng backup for rerun (optional)
• General recommendations:
• For most practical purposes 200-2000ng is enough
• Sample volumes: 10-30µl
• Sample concentration: preferably >400ng/ul to avoid speed vac. Not below 50ng/µl!
Low input RNA comes at the expense of reduced call rate
Workflow – ConsultationBiological 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 – ConsultationSample submission form
Customer sends filled in SSF to: [email protected]
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 – ConsultationSample submission form
Workflow – ConsultationSample submission form
Workflow – ConsultationSample submission form
Workflow – ConsultationSample 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 FedEx.
• 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)
OD260/280 > 1.7 (Nanodrop)
OD260/230 > 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 <1hr work is included in the price.
Follow up sales: microarray experiments lay the foundation for years of downstream work!
microarray services
PCR validation - services
Detection probes - ISH
Functional analysis – KD probes
In vivo KD
Systematic follow up on microarray service customers 2 – 4 weeks after final report
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Key selling points
• Exiqon miRCURY LNA™ microRNA microarrays• Superior sensitivity/specificity• Flexible for input amounts of total RNA 200ng – 2000ng• Works with FFPE – robust towards mRNA a rRNA degradation products• High coverage – miRBase 11 + 435 miRPlus• 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
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Thank you for your attention