The Facility provides automated DNA sequence analysis to the members of MUSC and unaffiliated researchers as well as other parties. Taq Dye Terminator chemistry is used for DNA sequencing. The service includes performing thermal cycling sequencing reactions on investigator-provided DNA templates followed by purification, polyacrylamide gel electrophoresis, laser-induced fluorescence detection analysis of the gel, data collection and analysis. Both plasmid DNA and PCR fragments can be sequenced either with custom primers, supplied by investigators, or any of the standard primers provided by the facility (T7 promoter, T7-terminator, M13 forward, M13 reverse, T3 and SP6). Sequence data are provided to investigators as four-color electropherogram plots and as text files delivered via E-mail or on a computer disc.
Free software is available from the web for viewing and editing the chromatogram output from our DNA sequencers. We recommend that our customers use these applications, since they can be helpful in resolving the ambiguities in the sequences and in trouble-shooting the problems.
The Facility’s applications are not limited to simple nucleotide sequence analysis. Microsatellite analysis, DNA fragment sizing for RFLP / AFLP analysis with the GeneScan software, as well as Single Nucleotide Polymorphism (SNP) analysis are also available. Variations in the DNA sequences of human genes can determine development and progression of diseases and responses to pathogens, chemicals, drugs and conventional therapies. Advances in the technologies of genetic analysis improved the detection of these variations (SNPs, insertions and deletions) and they are widely used for diagnosis of many diseases. The genotyping application of our instruments allows us to switch easily between microsatellite, SNP and general fragment analysis. The facility personnel can consult with investigators on the genomic analysis and assist in design of the approach for specific projects. On a case-by-case basis, the Nucleic Acid Analysis Facility will undertake genotyping projects on behalf of investigators, when this activity is defined in scope. Investigators should contact the NAA Facility Director for the information regarding Genetic Analysis.
The Nucleic Acid Analysis Facility also offers consultative aid in DNA template preparation and purification, and primer selection and design. Use of the primer analysis Software “Oligo 6.2” is available free of charge to all MUSC investigators