DNA Sequencing by Capillary Electrophoresis Using Short Oligonucleotide Primer Libraries

Abstract

Two strategies for DNA sequencing by primer walking using short oligonucleotide primer libraries have been successfully employed along with capillary electrophoresis using replaceable polymer solutions of linear polyacrylamide and fluorescence detection. A 3.5-kb stretch of the single-stranded M13mp18 template was sequenced with T7 PRISMTM dye-terminator/Sequenase® chemistry. An in-house base-calling program offered read lengths of roughly 450 bases with an average of 97.8% accuracy.