The Cleveland Clinic Genomics Core provides investigators (both academic and commercial) access to State-of-the-Art genomics capabilities: High Throughput Sequencing, Capillary Sequencing / Genotyping and Micro-arrays (Whole Genome Gene Expression, Genotyping and Methylation).
Contact: Mohammed Orloff, Ph.D orloffm@ccf.org
The Genomics Core is equipped with a 3730xl DNA analyzer and employs experienced, well-trained personnel to handle sequencing projects. However, a major part of sequencing success in achieved at the bench where the template DNA is prepared and its concentration is measured: properly purified DNA preparations give good sequencing results. Optimal results are dependent on the absence of excessive salts, organic solvents and residual ethanol in the template DNA preparation as well as the correct amount of template DNA. Qiagen products are preferred fro DNA isolations.
DNA samples for sequencing: Prepare DNA samples using the appropriate Qiagen product (recommended). Quantitate DNA and verify the amount and nature of the isolated DNA by running a diagnostic DNA digest out on an agarose gel.
Primers for sequencing: We recommend the following for primer length and content: length = 20-25 nucleotides, Tm = 56-75 °C, G-C content: 40-60%, combination of two GC’s at 3’ end.
The Genomics Core currently provides the following primers for DNA Sequencing at no charge to customers (These sequencing primers are available through the LRI storeroom in 50 µl aliquots at 5 pmol/µl which will facilitate submitting premixed template / primer solutions).
Sequencing difficult templates: For templates larger than 9 kb and known GC-rich templates we recommend our sequencing plus option which involves adding extra sequencing reagent. It greatly improves the success-rate of these reactions.
Continue to Genomics Core web site.