Mawi DNA Technologies LLC Next Generation BioSampling Devices Dr. Bassam El-Fahmawi President & CTO Collect – Stabilize – Concentrate – Transport – Extract – Store:  All in a Single Tube Mawi DNA Technologies LLC

About Mawi DNA Technologies Mawi DNA Technologies LLC: Established in Newark, California in 2012 iSWAB: 9 years of dedicated R&D from concept to commercialization 100% US-Made and Manufactured technology. Maryland California

Mawi DNA Technologies Mission Statement: Improving human and animal health assessment by enabling efficient and cost effective biological sample collection, room temperature stabilization and transport

Genomics Sample Collection Workflow Transport / Store Extract / Purify DNA Analyze

Invasive vs. Non-Invasive Collection Collect cell samples Stabilize sample in proprietary alcohol-free buffer Remove and discard swab Blood Painful Requires expertise Expensive handling infrastructure (cold chain transport and storage) iSWAB-DNA Collection Device Painless Enables self or assisted collection No cold chain involvement

Minimal storage foot print iSWAB vs. Blood iSWAB requires less collection time and no professional training ($80-$150) Vs. Self collection Requires specialized personnel iSWAB does not require cold transport Ambient shipment in standard mail Specialized & refrigerated transport iSWAB can be stored in supplied sample vial >1 year at room temperature Room Temperature storage vs. Refrigerated Specialized Storage Minimal storage foot print

iSWAB™ Collection Devices Vs iSWAB™ Collection Devices Vs. Other Non-Invasive Sample Collection Devices

Disadvantages of Traditional Non-Invasive Collection Methods Saliva/Mouthwash Oral Swabs Self collection, large volume of saliva required (1-2 mL) can take up to 45 mins limited to individuals capable of producing sufficient saliva (drugs that cause dry mouth as a side-effect can compromise collection) Collection solution contains alcohol (requires special extraction requirements and can possibly shear DNA) Yields sheared, short-fragmented DNA not optimal for complex downstream genomic applications Up to 40% bacterial content Readily available in any clinic, simple and easy to use Low gDNA recovery (up to 2 µg/Sample) with high bacterial contamination High resampling rate reduces operational efficiency and increases processing costs Transit time of dry swabs exponentially accelerates sample degradation and reduces DNA recovery substantially Swab removal and extraction process not automation-friendly

iSWAB vs. Saliva Collection iSWAB wins on Collection Time Swabs need to be purchased separately with children kits All collection accessories included iSWAB-DNA < 5 minutes Oragene iSWAB wins on Transportation- Bar-coding and Leak Safe Secure with O-ring Transport vessel included Bar-code comes standard for traceability No O-ring: Leakage Potential Transport vessel needed Bar-coding does not come standard iSWAB wins on Storage real-estate and cost 1/4th space needed vs. Oragene

iSWAB-DNA: Reinventing the swab Patent Pending Buccal cells collection and concentration device is bar-coded for traceability & chain of custody Innovative packaging designed to be tamper proof and a sample shipper box

Advantages of the iSWAB Non-Invasive Sample Collection Device Proprietary tube insert design and alcohol-free buffer yields up to 30mg of double-stranded, long fragment DNA ideal for complex downstream genomic applications iSWAB technology eliminates the need for swab retention Collects & concentrate predominantly buccal cells thus providing higher quality non-fragmented gDNA Less than 1% bacterial genomic DNA contamination results in pure sample DNA Self collection or assisted collection in less than 5 minutes. Suitable for all population segments including infants, toddlers, and elderly Swab-free sample transport decreases sample processing time without compromising sample integrity Long-term room temperature stability reduces sample storage and transport costs by eliminating cold chain requirements Manual and automation friendly tubes for scalability and cost efficiencies

Why Buccal Cells not Saliva? Buccal cells: best surrogate tissue for brain DNA methylation studies Buccal Cells Are the Dominant Cell Type in The ISWAB Saliva Content: Degraded cells WBCs High Bacteria Content Food particles Mucus nasal execrations iSWAB Content: Buccal Cells Low Bacteria Content Cell Types in ISWAB Post Collection: Total cell counts were assessed by a haemocytometer and differential cell counts by two observers from on Giemsa stained slides. The results are expressed a percentage of each cell type from the total number of cells in the iSWAB collected sample (n=10).

iSWAB-DNA Collection Process Accommodates extraction of cells from multiple swabs per patient Allows sample concentration Unique design eliminates the need for swab transport Self or assisted collection, accommodates all age groups and population segments Up to 30µg of double stranded, long-fragment human gDNA with <1% bacterial content 2-4 Minutes Collection iSWAB Sample Collection Process

iSWAB-DNA Collection Confirmation 4 Swabs

Lab Processing: Saliva vs. iSWAB Heat at 50C for an hour Specialized gDNA Extraction Procedures Compatible with any commercial Blood gDNA extraction kits with no modifications Faster sample processing ….. Faster TAT…….. $2-4$ saving per sample prep

Efficiency of iSWAB Sample Collection & Concentration Vs. Saliva Comparison of Average Yield of DNA from iSWAB-DNA & Oragene Saliva collection methods (OG-500 & OCR-100) Number of Samples = 48 Average OD 260/280 DNA Concentration (µg/Sample) iSWAB efficiently suspends and concentrate DNA from buccal cells iSWAB DNA yield and quality is comparable to Saliva and Superior over standard swab collection *DNA extraction was performed with DNeasy. DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used in QPCR to relatively quantitate the levels of Human genomic DNA

Low Bacterial Genomic DNA Contamination Comparison of Average Bacterial Genomic DNA Contamination Detected in Different Non-Invasive Sample Collection Methods Percentage of Bacterial Genomic DNA Content/ Prep iSWAB bacterial DNA content was determined by RT-PCR using primers specific for the 16S ribosomal RNA gene (n=24) The bacterial content from iSWAB was <1%, as compared to the other collections methods for the bacterial as reported in literature. iSWAB Proprietary cell lysis stabilizing solution allows for minimal to negligible bacterial DNA contamination

Collection Time and Age Affect on gDNA Yields From iSWAB

The Effect of the Sample Collection Time on DNA Yields  32%  32% DNA Concentration (µg/Sample) DNA extraction was performed with Gentra Puregene Blood extraction kit (n=6): Each volunteer was asked to self collect Buccal cells using the iSWAB device at different time intervals during the day. Volunteers where asked to abstain from eating, chewing gum and drinking for at least 30 minutes before colleting the samples. Early morning sample was collected first thing in the day and before brushing their teeth. Yields where lower by 32% in midday and 20% in the evening from early morning collections. *DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used to relatively quantitate the levels of Human genomic DNA .

ISWAB is an Efficient Non-Invasive Sample Collection for Various Population Segments DNA Concentration (µg/Sample) DNA extraction was performed with Gentra Puregene Blood extraction kit (n=34/age group): Age groups over 14 years old has performed self collection, where the collection for younger age groups were performed by one of the parents. Sample collection was performed regardless of the time of the day. *DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used to relatively quantitate the levels of Human genomic DNA

iSWAB/Saliva/Blood Sample Comparison Objectives: Customer was looking for a convenient  DNA collection device that could be easily shipped without refrigeration and be used in clinical DNA sequencing Conclusions: iSWAB DNA performed As Well or Better than blood and better than saliva Sequence analysis of all samples revealed no significant difference between sample types iSWAB extracted DNA is usable for carrier screening assay The proportional increase of PCR sensitivity to the gDNA input from iSWAB-DNA device is an indication of very limited PCR inhibitor carryover

iSWAB-DNA Performance in NGS: MiSeq Run FastQ High Alignment, Minimal bacterial DNA Contamination

iSWAB-: Collecting and Preserving DNA Evidence at the Point of Collection Touch DNA & Reference Samples

ISWAB-ID Compatibility with Direct PCR- ID Profiling Kits ABI Identiflier Direct PCR STR kit

ISWAB-Blood for the Blood Drops Collection Collect – Concentrate – Transport – Store:  All in a Single Tube Launch Timeline: ASHG – Oct 7th, 2015 MAWI DNA Technologies LLC

Current Blood Collection Methods Venous Blood Dry Blood Spot Painful (invasive) Large volume blood collected (5-10 mL) Requires expertise Expensive handling infrastructure (cold chain transport and storage High gDNA yields with >23 Kb fragment sizes Finger brick (minimally invasive) Five blood spot collected or more Proper collection requires the blood drop to go through both sides of the paper within designed circle Three hours dry time before transport or storage Low yields and highly fragmented DNA

ISWAB- Blood Collection Process Two Sterile Cotton Swabs Finger disinfectant pad Lancet to obtain blood from your finger Carrying case for Lancet and its accessories ISWAB-Blood Tube ISWAB-Blood Tube Rack of 50 ISWAB-Blood Collection Instruction Video

ISWAB-Blood Vs. Dry Spot Cards

High Molecular Weight gDNA From ISWAB-Blood Spots Agarose gel (0.8%) electrophoresis of gDNA samples isolated from 100 µL of Human blood collected with ISWAB-Blood Spot using QiaAMP Blood kit. 4 µL of 100 µL elute was used for electrophoresis. M:λDNA/Hind III+EcoR I

Real Time Stability Testing at Room Temperature: kb 23.1 9.4 6.5 4.3 2.3 2.0 0- days 7- days 30- days 45- days 180- days DNA extraction was performed with QiaAMP Blood extraction kit Pooled ISWAB-Blood samples (n=24) was subjected to actual non-refrigerated airfreight sample shipping conditions, then stored at room temperature. An aliquate of 200 µL from the pooled sample was processed at each time point with QiaAMP Blood mini (whole blood protocol), then 2 µL of 100 µL elute was used for electrophoresis. Human HMW gDNA integrity is maintained and nuclease activity neutralized over the real time testing period of 6 months

Long Term Stabilization of Blood Analytes

ISWAB-Blood Advantage No dry time required, 5 minutes collection process After collection, the sample ius ready for room temperature shipment and long term storage Unlike FTA or FTA like cards, extracting DNA or measuring blood analytes does not require any front prep and/or card punching DNA extraction can be performed with standard whole blood purification protocol (2- 8 µg of high MW gDNA) Easy access to the sample for screening and/or in depth analysis purposes (genomics & metabolmics) – reducing resampling and assay failures Manual and automation compatible with no pre-prep process

iSWAB Automation Compatibility

ISWAB-DNA Genomics Workflow Compatibility iSWAB has been successfully tested on various genetic analysis platforms for human, animal, or forensic samples iSWAB-DNA No DNA Purification Required DNA purification recommended - Extraction can be performed using the “whole blood” protocol of most commercially available kits qPCR Sanger Sequencing NGS Direct PCR & RT-PCR Microarrays

DNA from iSWAB is Ideal for Simple & Complex Downstream Analysis

iSWAB Customizable Solutions We offer a number of flexible options for customization Labeling and Packaging Your branded labels on our box and/or collection devices Customized return address labels Include your forms in our package Custom barcodes Custom Tube Components Our patented squeezing insert can be custom molded to fit your vial of choice, and accommodate any size swab head. We can accommodate different caps including pierceable caps for automation or specific decapper-compatible. For inquiries please contact us at mawidna.com/distributors

ISWAB Product Offering iSWAB-DNA iSWAB-RNA iSWAB-Protein (For drugs screening and protein biomarkers) iSWAB-Vet / Animals iSWAB-ID (Foresncis and Paternity testing) Coming Soon Oct 7th, 2015 iSWAB-Blood (for collecting blood drops from newborn babies) Q1-2016 iSWAB-CELLS (Collecting and transporting viable cells up to 4 weeks at RT) – Regenerative medicine, metagenomics, OMICS, Microbiome , & Cell lines transport Q2-2016 MRT-Blood (Whole blood transport with 15 days stability)

Reinventing Non-Invasive Sampling! Thank you!