Open AccessDNA Sequencing by Capillary Electrophoresis Using Short Oligonucleotide Primer Libraries
Author(s) -
Marie C. RuizMartinez,
Emanuel Carrilho,
Jan Berka,
Jan Kieleczawa,
Arthur W. Miller,
František Foret,
Spencer Carson,
Barry L. Karger
Publication year - 1996
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/96206rr03
Subject(s) - capillary electrophoresis , oligonucleotide , primer (cosmetics) , dna , terminator (solar) , microbiology and biotechnology , dna sequencing , dna sequencer , biology , electrophoresis , gel electrophoresis , base pair , genetics , computational biology , chemistry , ionosphere , physics , organic chemistry , astronomy
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 PRISM TM dye-terminator/Sequenase ® chemistry. An in-house base-calling program offered read lengths of roughly 450 bases with an average of 97.8% accuracy.