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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
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

you re collecting patient samples now what
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
they are still your samples
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.
dna extraction from blood
DNA extraction from blood
  • Gentra Puregene reagent system
  • ~350 µg DNA from 10 ml blood
  • Autopure machine
  • Bar coded
dna archiving
DNA archiving
  • DNA tubes: ID #, bar coded
  • Puregene reagent system:

Purified DNA is stored dissolved in TE at 4˚. Stable for >10 years

dna extraction standard service
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.
additional services
Additional services
  • Lymphoblast immortalization & cell line banking
  • Whole-genome amplification for depleted DNAs
  • Real-time PCR
genotyping services
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

genotyping services9
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

genotyping services10
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).

the dna resources core 518 light hall11
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