Simplified template preparation and improved direct sequencing using Taq polymerase.
A streamlined version of direct dideoxy sequencing is presented that includes template preparation as well as sequencing protocols. The method is used routinely to sequence double-stranded PCR products after minimal purification with one of the primers used in amplification. Either 35S or 32P labeling can be used with equally good results.
Srivastava
A K
AK
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110.
Montanaro
V
V
Kere
J
J
D004259:DNA-Directed DNA Polymerase; D004587:Electrophoresis, Agar Gel; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D019914:Taq Polymerase; D013698:Templates, Genetic
D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase