Transcript The DNA Resources Core 518 Light Hall • Manager: Cara Sutcliffe 6-2744

The DNA Resources Core
518 Light Hall
• Manager: Cara Sutcliffe
6-2744
[email protected]
• Faculty Director: Doug Mortlock
6-1671
[email protected]
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
• 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.
They are still your samples!
DNA extraction from blood
• Gentra Puregene
reagent system
• ~350 µg DNA from
10 ml blood
• Autopure machine
• Bar coded
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 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
• Lymphoblast immortalization & cell line
banking
• Whole-genome amplification for depleted
DNAs
• Real-time PCR
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 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 services
• Service models using the ABI 7900HT
genotyping platform:
1. “Full service”: Customer selects Assays-ondemand; 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 Hall
• Manager: Cara Sutcliffe
6-2744
[email protected]
• Faculty Director: Doug Mortlock
6-1671
[email protected]