Abstract: A novel method of primer walking using octamer oligonucleotides to prime DNA sequencing reactions is described. Octamer sequencing is compatible with isotopic and fluorescent sequencing chemistry, reaction conditions are optimized such that the samples can be processed in parallel and the procedures are automated. This strategy is faster than the traditional primer walking sequencing strategy as the existence of a primer library allows immediate access to a primer for the next sequencing reaction, eliminating delays associated with designing and synthesizing gene specific primers. The octamer library is comprised of optimized sequencing primers, such that octamer sequencing yields results equivalent to or better than traditional primer walking.

Abstract: Guanine-rich oligonucleotide primers and short telomeric DNA oligonucleotides that serve as non-conventional templates for the synthesis of long, nascent DNA strands are used to screen for pharmaceutical agents. Using the methods of the invention, agents exhibiting antagonistic or agonistic effects on guanine quartet formation can be selected for further study. The invention also provides a predictive assay for cancer in which the level of DNA polymerase activity at a quartet stabilized template is measured. The invention also contributes a method for screening pharmaceutical agents that reduce or inhibit DNA polymerase activity at guanine quartet stabilized templates. The invention provides a method of creating extended DNA molecules from oligonucleotide templates. The invention also provides a method of inhibiting the ageing process in which an oligonucleotide capable of forming non-Watson-Crick structures is used to promote the elongation of telomeres by DNA polymerase.