the dna resources core 518 light hall

The DNA Resources Core518 Light Hall

• Manager: Cara Sutcliffe

6-2744cara@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 • 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!

• 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• 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 assaysTypically >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 assaysTypically >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-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 Core518 Light Hall

• Manager: Cara Sutcliffe

6-2744cara@chgr.mc.vanderbilt.edu

• Faculty Director: Doug Mortlock

6-1671

mortlock@chgr.mc.vanderbilt.edu