Principles of Capillary-Based Sequencing for Clinical Microbiologists

Abstract

Chain termination cycle sequencing, or “first-generation” DNA sequencing, was developed 3 decades ago but remains one of the most commonly used procedures for diagnostic analyses. Automated capillary-gel electrophoresis genetic analyzers greatly improved the efficiency of sequencing DNA templates between 100 and approximately 1,300 nucleotides long. Cycle sequencing may be completed the same day by using fast protocols for the initial amplification and cycle-sequencing reactions and by utilization of commercial sequence interpretation and analysis software. These changes allowed sequencing to become a routine tool for pathogen identification, discovery, and genotyping.