| Abstract
| - Genotyping based on short tandem repeat (STR) regionsis used in human identification and parentage testing,gene mapping studies, cancer diagnostics, and diagnosisof hereditary diseases. Analysis of STR systems using slabgel electrophoresis requires lengthy and labor-intensiveprocedures. Therefore, alternative methods such as capillary electrophoresis or ion-pair reversed-phase high-performance liquid chromatography (IPRP HPLC) havebeen used to analyze DNA. IPRP HPLC offers an attractivesubstitute to gel electrophoresis for STR analysis becauseof the reduced analysis time, and there is no need for thewaste disposal associated with radioisotopic, enzyme-linked, or fluorescence detection systems. We evaluatedthe use of IPRP HPLC for the sizing and typing of STRalleles from the HUMTHOl locus. The IPRP HPLC conditions (column temperature, flow rate, percent organicmodifier per minute) were optimized for the separationof PCR products. Using the optimized separation conditions, the alleles of the HUMTHO1 system were sized intheir native state (double stranded) with the use of internalmarkers. The typing results correlated 100% to acceptedmethods of DNA typing. The analysis time for theHUMTHOl locus was less than 14 min, and the allelescould be peak captured for further examination followingsuch as sequencing.
|
