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Dry State, Room Temperature, BioSample Management Solutions

Dry State, Room Temperature, BioSample Management Solutions For the collection, transport, storage and recovery of genetic material. standardizing biosample management. simplifying workflows. enabling scientific discoveries. Tissue Bank Mini-Symposium

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Dry State, Room Temperature, BioSample Management Solutions

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  1. Dry State, Room Temperature, BioSample Management Solutions For the collection, transport, storage and recovery of genetic material standardizing biosample management. simplifying workflows. enabling scientific discoveries. Tissue Bank Mini-Symposium Presented by: David Wellis, Ph.D, CEO December 4, 2008

  2. The Biomedical Research EcoSystem

  3. Cryostorage – Current State of Art in Sample Management • Inefficient aliquot usage (freeze/thaw) • Manual operation, usually • Energy intensive • Space consuming • Overnight hazardous transport

  4. GenVault’s Dry State Solution • Automation-friendly • Guaranteed sample preservation with minimal sample waste • Dry state format optimizes sample quality and requires minimal energy • High-density GenPlates hold 10-14x the number of aliquots in the same space as freezers • Bone-dry GenTegra integrates seamlessly into most workflows GenPlates/GenTegra for aliquots Archive for storage GenSolve for GenPlate DNA recovery GenConnect software

  5. How does it work? The GenPlate enables room temperature storage by incorporating FTA paper • Proven DNA stabilizer with 18 years of data • Lyses cells upon contact • Inactivates viruses and bacteria • Ship at room temperature with no biohazard labeling FTA: Chemically treated porous Paper allows for DNA storage at room temperature. - Courtesy of Whatman

  6. GenSolve for DNA Recovery DNA recovered from blood spotted on FTA paper using GenSolve can be… (1) …amplified reliably (2)…at least 35kb without fragmenting Simple DNA Recovery process with few hands-on steps Recovered DNA is double-stranded allowing for usage in a wider variety of downstream applications, particularly genotyping

  7. GenPlates for Various Sample Types and Storage Densities ~ 150 ng DNA per element 1 or 3 Region GenPlate Blood ~ 600 ng DNA per element 6 Region GenPlate Buffy Coat ~ 500 ng to 3µg DNA per element 12 Region GenPlate Purified DNA ~ 80-90% recovery of purified plasmids or glycerol and DMSO stocks: 24 Region GenPlate Clones, Plasmids ~ 500 ng DNA per element 6 or 12 Region GenPlate Buccal MW ~ 25 ng DNA per element 24 Region GenPlate Buccal Swab Varied formats optimized for sample types and applications

  8. Fresh Tissue Application Proof of Principle Data

  9. Preservation of Fresh Tissue on GenPlates • With GenVault, a process for digesting fresh tissue and then spotting the digest onto GenPlates was tested: • Begin with ~25mg of tissue in small pieces • Add Qiagen Protease and Buffer ATL then incubate at 56°C for 1-3 hours • Vortex the sample vigorously • Apply 10µl to each well in a GenPlate

  10. DNA Recovery from GenPlates Step 1: Place elements into a tube Step 2: Incubate with GenSolve reagents for 1 hour Step 3: Centrifuge Step 4: Proceed with DNA purification

  11. Recovery of DNA from Fresh Tissue Spotted GenPlates Four leiomyoma samples were digested and spotted onto GenPlates in masses ranging from 7mg to 13mg Two of these samples were stored in RNA Later prior to spotting and two were stored in RPMI media Elements were recovered in pools of 1, 3 or 6. Yields linearly increased with additional elements Yields from 3 and 6 element elutions were suitable for high-throughput genotyping assays

  12. 0.8% E Gel Run for 30 min Quality of Recovered DNA An aliquot of each of the sample types above was run for 30 minutes on a 0.8% E-gel (Invitrogen) to determine DNA length and to verify quantitation. 50 ng of each sample per Picogreen determination was run in each lane. Lane 1 = Apex ladder with the top band at 35kb Lane 2 = Roche genomic DNA Lane 3 = whole blood sample Lane 4 = buffy sample Lane 5 = tissue RPMI #1 Lane 6 = tissue RPMI #2 Lane 7 = tissue RNA Later #3 Lane 8 = tissue RNA later #4 1 2 3 4 5 6 7 8 35 kb All 6 sample types tested appear to be full length (35kb) DNA from fresh tissue is comparable to DNA extracted from whole blood or buffy coat

  13. Formalin-Fixed, Paraffin-Embedded (FFPE) A Novel Approach for the Recovery of DNA from a Difficult Source

  14. Preservation of FFPE Tissues on GenPlates • Archival tissue samples are an important source of DNA • Recovery of DNA from these formalin-fixed, paraffin-embedded (FFPE) tissues has historically been extremely difficult. • Low content of DNA • Existing DNA is poor quality • Contaminants from the FFPE fixing process are difficult to remove and impede further analysis • Additionally results can vary from slice to slice • GenVault has developed a process which liquifies FFPE tissue slices and then stabilizes them in a standardized, consistent format: GenPlates • Dry-state, room temperature storage • Standard storage format • Consistent delivery of DNA suitable for many types of genetic analysis

  15. FFPE Liquification and Storage Process • Step 1: Place tissue slice into BiOstic Paraffin Removal Reagent to de-paraffinize slices • Step 2: Wash tissue slice with 100% EtOH • Step 3: Lyse samples • Step 4: Spot liquid in 10µl aliquots onto GenPlate elements

  16. DNA Recovery from GenPlates Step 1: Place elements into a tube Step 2: Incubate with GenSolve reagents for 1 hour Step 3: Centrifuge Step 4: Proceed with DNA purification

  17. DNA Quality from GenPlates DNA recovered from FFPE tissue spotted GenPlates was run on a 0.8% agarose gel The majority of fragments are less than 1kb This size range is sufficient for many types of genetic analysis such as whole genome amplification, PCR and many types of genotyping assays

  18. DNA Yields from GenPlates • Yields may vary from tissue type to tissue type • DNA recovered after storage on GenPlates is comparable to direct purification from tissue slices • Yield is sufficient for many types of genetic analysis

  19. DNA Quality from FFPE DNA recovered from FFPE spotted GenPlate elements was used as template in qPCR Compared to Roche DNA controls, and direct purification, the DNA recovered by this process shows no PCR inhibition

  20. Carle Foundation –Tissue Application in Use

  21. Carle Foundation: Tissue Application in Use

  22. Summary GenVault’s dry state, room temperature biosample management solutions provide high quality, consistent results, for a variety of starting material, including fresh and FFPE tissue samples.

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