"First generation" automated DNA sequencing technology.
ABSTRACT Beginning in the 1980s, automation of DNA sequencing has greatly increased throughput, reduced costs, and enabled large projects to be completed more easily. The development of automation technology paralleled the development of other aspects of DNA sequencing: better enzymes and chemistry, separation and imaging technology, sequencing protocols, robotics, and computational advancements (including base-calling algorithms with quality scores, database developments, and sequence analysis programs). Despite the emergence of high-throughput sequencing platforms, automated Sanger sequencing technology remains useful for many applications. This unit provides background and a description of the "First-Generation" automated DNA sequencing technology. It also includes protocols for using the current Applied Biosystems (ABI) automated DNA sequencing machines.
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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.Clinical Microbiology Newsletter 01/2013; 35(2):11–18.