chapter  19
7 Pages

Octamer-Primed Cycle Sequencing

We have developed a primer-directed DNA sequencing method (octamer sequencing technology [OST]) in which individual octamers are selected from a presynthesized primer library and used to prime multiple DNA sequencing reactions in parallel.1-3 The availability of such a library eliminates delays associated with designing and synthesizing gene-specific primers. Furthermore, the primers in our working octamer library can be used to sequence virtually any template. A computer program, electronic OST (eOST), was developed to identify the optimal octamer-primers for a particular template

and to facilitate the process of primer selection.4 In addition, we discovered that nucleotide supplementation of octamerprimed sequencing reactions leads to an increase in the success rate of fluorescent sequencing and improves the quality of the sequence data obtained from difficult template regions.5