OCTAMER-PRIMED CYCLE SEQUENCING - DESIGN OF AN OPTIMIZED PRIMER LIBRARY

This paper describes a novel method of primer walking using octamer ol igonucleotides to prime DNA sequencing reactions. Octamer sequencing i s compatible with isotopic and fluorescent sequencing chemistry, react ion conditions are optimized such that the samples can be processed in parallel, and the procedure has the potential to be automated. This s trategy is faster than the traditional primer walking sequencing strat egy, as the existence of a primer library allows immediate access to a primer for the next sequencing reaction, eliminating delays associate d with designing and synthesizing gene-specific primers. The octamer l ibrary is comprised of optimized sequencing primers, such that octamer sequencing yields results equivalent to or better than traditional pr imer walking. This technology is more economical because gene-specific sequencing primers, the major cost in the reaction, are replaced by a n optimized subset of frequently occurring octamers that are able to p rime multiple reactions.