DNA Sequencing: An Outsourcing Guide

DNA sequencing, the process of determining the precise order of nucleotides in a DNA strand, is a fundamental and ubiquitous method in molecular biology. Examples of its many uses include characterization of unknown DNA, mutation detection, allele identification, and clone construct confirmation. Advances in sequencing technology, motivated by the successful International Human Genome Project, have resulted in unprecedented access to high-quality and cost-effective automated analysis. It is increasingly common for researchers to delegate routine DNA sequencing needs to dedicated laboratories or companies specializing in this technology. In this unit, we discuss key protocols for maximizing the success of DNA sequencing when working with such service providers and describe detailed troubleshooting advice for analyzing anomalous results. Keywords: DNA sequencing; capillary electrophoresis; PCR cleanup; plasmid preparation; chromatogram