The oligos are provided in autoclaved distilled water, ready for use for most applications without further purification. Concentrations are determined by UV absorbance at 260 nanometers and a custom report providing useful information such as the melting temperature, molecular weight, potential secondary structure, and a conversion factor for determining molarity is provided with every oligo prepared in the facility. The facility also offers oligo analysis using an Applied Biosystems Model 600 Capillary Gel Electrophoresis System. The DNA Facility also offers DNA sequencing on a fee for service basis. Sequencing is performed using fluorescent dye terminator chemistry and analyzed on an Applied Biosystems 373 Automated DNA Sequencer.
Investigators provide the facility with the DNA templates for analysis.
Reactions are performed using either standard sequencing primers provided by the facility or custom sequencing primers provided by the investigator. Most sequencing reactions employ the use of Taq DNA polymerase and a cycle sequencing protocol. The use of four different fluorescent dye dideoxynucleotide terminators permits single lane analysis for each template. Quality templates generate 350-500 bases of sequence information per reaction. Automatic base-calling software converts the raw date to sequence information.