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The DNA Resources Core 518 Light Hall

The DNA Resources Core 518 Light Hall. Manager: Cara Sutcliffe 6-2744 cara@chgr.mc.vanderbilt.edu Faculty Director: Doug Mortlock 6-1671 mortlock@chgr.mc.vanderbilt.edu. You’re collecting patient samples …Now what?. DNA extraction from blood or buccal swabs Storage of extracted DNA

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The DNA Resources Core 518 Light Hall

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  1. The DNA Resources Core518 Light Hall • Manager: Cara Sutcliffe 6-2744 cara@chgr.mc.vanderbilt.edu • Faculty Director: Doug Mortlock 6-1671 mortlock@chgr.mc.vanderbilt.edu

  2. You’re collecting patient samples …Now what? • DNA extraction from blood or buccal swabs • Storage of extracted DNA • Secure, but practical database system for managing samples • DNA retrieval at a later time, for genotyping samples of interest • SNP genotyping service appropriate to your scale and needs • Compliant with HIPAA

  3. They are still your samples! • Control of samples belongs to the investigator • Password-protected, Web-based database • Non-authorized personnel cannot access sample information or place requisitions for DNA aliquots • “Outsourcing” extraction, storage and organization of DNA samples.

  4. DNA extraction from blood • Gentra Puregene reagent system • ~350 µg DNA from 10 ml blood • Autopure machine • Bar coded

  5. DNA archiving • DNA tubes: ID #, bar coded • Puregene reagent system: Purified DNA is stored dissolved in TE at 4˚. Stable for >10 years

  6. DNA extraction: standard service • DNA Extraction from Whole Blood For 10mls (or 20mls) of blood, services include: • logging in and maintenance of sample information in our database, • DNA extraction, • archiving backup blood spot cards and frozen blood aliquots, • DNA quantitation by fluorometry, • one DNA requisition, • storage of extracted DNA for 5 years.

  7. Additional services • Lymphoblast immortalization & cell line banking • Whole-genome amplification for depleted DNAs • Real-time PCR

  8. Genotyping services • ABI 7900HT machine (Dan Roden/Pharmacogenetics of Arrythmia Therapy) • TaqMan SNP assays Typically >99% accurate • Assays-on-demand: Pre-validated SNP assays by ABI • Assays-by-design: For “custom” SNPs; need to be validated before use

  9. Genotyping services • ABI 7900HT machine (Dan Roden/Pharmacogenetics of Arrythmia Therapy) • TaqMan SNP assays Typically >99% accurate • Assays-on-demand: Pre-validated SNP assays by ABI • Assays-by-design: For “custom” SNPs; need to be validated before use

  10. Genotyping services • Service models using the ABI 7900HT genotyping platform: 1. “Full service”: Customer selects Assays-on-demand; core staff set up TaqMan PCR assays in 384 well plates, and scan plates on 7900HT to collect genotypes. ($400 setup + $0.80/genotype) 2. As 1, and core staff assist with designing custom Assays-by-design (inquire) 3. Customer obtains Assays-by-design; core staff perform PCR and scan plates. ($95 setup + $0.80/genotype) 4. Customer is responsible for setting up PCR and scanning plates ($10/plate).

  11. The DNA Resources Core518 Light Hall • Manager: Cara Sutcliffe 6-2744 cara@chgr.mc.vanderbilt.edu • Faculty Director: Doug Mortlock 6-1671 mortlock@chgr.mc.vanderbilt.edu

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