Journal Article

Application of N-terminally Truncated DNA Polymerase from <i>Thermus thermophilus</i> (Δ<i>Tth</i> Polymerase) to DNA Sequencing and Polymerase Chain Reactions: Comparative Study of Δ<i>Tth</i> and Wild-Type <i>Tth</i> Polymerases

Taku Arakawa, Boonsri Jongsareejit, Yusaku Tatsumi, Keiko Tanaka, Katsunori Ikeda, Hideyuki Komatsubara, Hiroaki Inoue, Bunsei Kawakami, Masanori Oka, Shigenori Emi, Tetsuya Yomo, Yasufumi Shima, Seiji Negoro and Itaru Urabe

in DNA Research

Published on behalf of Kazusa DNA Research Institute

Volume 3, issue 2, pages 87-92
Published in print January 1996 | ISSN: 1340-2838
Published online January 1996 | e-ISSN: 1756-1663 | DOI: http://dx.doi.org/10.1093/dnares/3.2.87

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N-Terminally truncated DNA polymerase from Thermus thermophilusTth polymerase) lacking 5′-3′ exonuclease activity was used for DNA sequencing and polymerase chain reaction (PCR). In contrast to the high background of the sequencing ladder observed with the wild-type Tth polymerase, ΔTth polymerase gave readable sequencing patterns which extend up to more than 500 bases from the primer site on cycle sequencing and automated sequencing. The ΔTth polymerase was used for the standard and mutagenic PCR, and net amplification of the DNA and the mutations accumulated during PCR were analyzed. Under mutagenic PCR, the mutation rates were 7.0 × 10−4 (Tth) and 8.3 × 10−4Tth) per nucleotide per cycle of amplification, which were 4–9 times higher than the rates under standard PCR.

Keywords: Thermostable DNA polymerase; Thermus thermophilus; Polymerase chain reaction

Journal Article.  0 words. 

Subjects: Genetics and Genomics

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