the dna resources core 518 light hall manager: cara sutcliffe 6-2744 [email protected]...
TRANSCRIPT
![Page 1: 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](https://reader035.vdocuments.us/reader035/viewer/2022062722/56649f285503460f94c408ee/html5/thumbnails/1.jpg)
The DNA Resources Core518 Light Hall
• Manager: Cara Sutcliffe
• Faculty Director: Doug Mortlock
6-1671
![Page 2: 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](https://reader035.vdocuments.us/reader035/viewer/2022062722/56649f285503460f94c408ee/html5/thumbnails/2.jpg)
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
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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.
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DNA extraction from blood• Gentra Puregene
reagent system
• ~350 µg DNA from 10 ml blood
• Autopure machine
• Bar coded
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DNA archiving
• DNA tubes: ID #, bar coded
• Puregene reagent system:
Purified DNA is stored dissolved in TE at 4˚. Stable for >10 years
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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.
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Additional services
• Lymphoblast immortalization & cell line banking
• Whole-genome amplification for depleted DNAs
• Real-time PCR
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Genotyping services
• ABI 7900HT machine(Dan Roden/Pharmacogenetics of Arrythmia Therapy)
• TaqMan SNP assaysTypically >99% accurate• Assays-on-demand: Pre-validated SNP assays by ABI• Assays-by-design:For “custom” SNPs; need to be
validated before use
![Page 9: 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](https://reader035.vdocuments.us/reader035/viewer/2022062722/56649f285503460f94c408ee/html5/thumbnails/9.jpg)
Genotyping services
• ABI 7900HT machine(Dan Roden/Pharmacogenetics of Arrythmia Therapy)
• TaqMan SNP assaysTypically >99% accurate• Assays-on-demand: Pre-validated SNP assays by ABI• Assays-by-design:For “custom” SNPs; need to be
validated before use
![Page 10: 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](https://reader035.vdocuments.us/reader035/viewer/2022062722/56649f285503460f94c408ee/html5/thumbnails/10.jpg)
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).
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The DNA Resources Core518 Light Hall
• Manager: Cara Sutcliffe
• Faculty Director: Doug Mortlock
6-1671