Automated sequencing technology

The strategy described below involves direct cycle sequencing of the PCR product, using the Dye Terminator technology for an ABI automated sequencer (ABI PRISM Dye Terminator Cycle Sequencing Core Kit, Perkin-Elmer). 

Non-SELEX is an attractive alternative to SELEX, due to increased speed and suitability for easy automation. Non-SELEX leads to a final pool with all sequences being unique. The analysis of such sequences would be much more efficient if single-molecule sequencing were used. In this respect, the current effort in the development of single-molecule sequencing technologies is quite encouraging (Bayley, 2006). 

The chemistry and instrumentation described above were adequate to demonstrate the feasibility of automated sequencing using multiple fluorophores. However, for this technology to be widely applicable it was necessary for it to be commercialized. Accordingly, we undertook a collaboration with Applied Biosystems Inc. (ABI) to develop a commercial version of the instrument. Several significant changes to the technology were made (6). 

In summary, the current instrument for automated DNA sequencing, although a novel and powerful new tool, represents just a first step in the development of the technology. As significant improvements in the various aspects of these methods are achieved, automated sequencing will become an increasingly powerful tool for use in modern medical and biological research. 

It is likely that alternative sequencing technology will displace electrophoretic sequencing. Microfabricated capillary array electrophoresis has been studied for human identification via STRs. Genetic analysis by PCR has been reported for prenatal diagnosis of beta-thalassemia. Microchip devices can automate the complete DNA analytical process, including DNA extraction, cleanup, PCR, and size separation." ... 

This sequence of steps, shown schematically in Figure 1, can be repeated many times in the course of an automation effort, as new needs arise from emerging product and process technologies and from experimental results. 

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