DNA Sequencing and Gene Analysis Center
In addition to automated DNA sequencing, the Center specializes in genotype analysis of drug metabolizing enzymes such as Cytochrome P450 (isozymes CYP2D6, CYP2C19, and CYP2C9), and drug transporters such as products of multidrug resistance genes (MDRs, MRPs). We also offer Realtime® PCR testing technology for SNP genotyping and quantitative RT-PCR analyses. Please contact Dr. Edward Kelly, the center manager at 616-8718 for additional information on genotype and of RT-PCR analysis projects.
Description of DNA sequencing service:
The Center accepts DNA samples (pure plasmids, mini-prep plasmids, M13 clones, cosmids, lambda clones, PCR products, gel isolated fragments) and processes them according to protocols for Applied Biosystems DNA Sequencers. One sequencing run (one template, one primer) typically produces greater than 600 nucleotides of high-quality sequence data. The results are returned to the researcher as chromatogram (adobe PDF or postscript files), and text sequence files. Investigators must supply a CD or flash memory stick for this purpose. On request and for a nominal fee, we can also print the data, and place it in a pickup bin outside the door.Quality of data:
The sequence returned to investigators is usually 600-700 nucleotides long. The reliable length of sequence data depends on a variety of factors. A good quality of template DNA and the primer will ensure maximum length and reliability of sequence data. It is crucial for each customer to evaluate their own data and to determine the extent of reliable data. Because data evaluation is highly dependent on your individual needs, we cannot perform this step for you.
Therefore, we provide the Sequencing Chromatograms in PDF or PS files format for further examination according to your research need. If you are unsure how to evaluate these files, please consult our staff.
The Center has no control over the quality of template or the design of the primer. However, we can and will do our best to help you to avoid such problems based on our collective experience. Nevertheless, we have to charge for sequencing reactions that fail for reasons outside of our control. We can provide validation data for control sample tested along with your submitted DNA samples.
Typical problems encountered:
Automated sequencing is very reliable in the hands of our experienced technicians. As a precaution, we always check that control samples sequenced properly, that excess terminators were removed, that the resolution was adequate and that the computerized data extraction proceeded normally. If a problem is detected, your samples are repeated at our cost. Each run is internally validated to ensure that reaction chemistry as well as separation steps are working properly before the results are sent back to our clients.
- Real Time PCR example
- Allelic Discrimination/SNP Genotyping example
- Sequencing Electropherogram example (PDF File)
How to Contact Us:
Please call Dr. Kelly at (206) 685-4641, or e-mail firstname.lastname@example.org for additional information on your project needs. For projects and grant proposals requiring DNA sequencing or genotyping analyses, please contact the Center Director Rodney Ho at (206) 543-9434.