Services & Protocols
The center offers a wide range of services from tissue to microarray and analysis.
Microarrays
We process Affymetrix and standard glass slide microarrays. Because microarrays are such an individualized procedure, please call us to discuss your specific needs.
Real-Time Quantitative PCR
The Bio-Rad iCycler Real Time PCR Detection System is used for real-time quantization of DNA. If necessary, the staff will help create templates by reverse transcription, set up reactions, or run plates that have already been set up. Data can be e-mailed to you or saved to CD or flash drive.
Gel and Blot Imaging
The GE Healthcare Typhoon 9410 is a high-performance gel and blot imager with the capability of microarray analysis. It offers three methods of detection: storage phosphor, fluorescence, and chemiluminescence. This system handles gel sandwiches, agarose and polyacrylamide gels, membranes, microplates, and microarrays. Red, green, and blue excitation wavelengths and a wide choice of emission filters enable imaging of an extensive variety of fluorophores. Automated four-color fluorescence scanning allows multiplexing of multiple targets in the same sample.
DNA Purification
Bacterial plasmids are purified in a 96-well format using a Qiagen 3000 BioRobot. We use the Qiagen R.E.A.L. prep. We can purify fewer than 96 samples if required.
DNA Quantitation
Double-stranded DNA is quantified using a fluorescent nucleic acid stain (PicoGreen®) and read on a Labsystems Fluoroskan Ascent fluorescence plate reader. The commonly used technique for measuring nucleic acid concentration by the absorbance at 260 nm (A260) has the disadvantage of being affected by the presence of nucleotides, single-stranded nucleic acids, and proteins — all contaminants commonly found in DNA preparations. However, PicoGreen® measures only the double-stranded DNA present.
PCR Cleanup
We use the Qiagen MinElute filter plate on the Qiagen BioRobot 3000 for PCR cleanup. This is a size exclusion membrane, so if your PCR reactions yield more than one product, they will all be purified. In order to successfully sequence PCR products that have been purified with the filter plate, either there must be only one PCR product in the sample or the sample must be sequenced with an internal primer that is specific to the product of interest. We can process any number of samples.
Real-Time Quantitative PCR
The ABI Prism 7000 Sequence Detection System is used for real-time quantization of DNA. Currently, the NGC does not create templates by reverse transcription or set up reactions. The instrument is available to run plates that have already been set up. Your data will be e-mailed to you in the form of an Applied Biosystems .sds file.