Accuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase

Accuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase

Autor Diaz, Ricardo Sobhie Autor UNIFESP Google Scholar
Sabino, Ester Cerdeira Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Irwin Memorial Blood Centers
Fundação Pró-Sangue, Hemocentro de São Paulo
Resumo For certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading activity, lacking in Taq polymerase, would be an improvement when higher fidelity is needed. We used the forward mutation assay to compare the fidelity of Taq polymerase and Thermotoga maritima (ULTMA ) DNA polymerase, an enzyme that does have proofreading activity. We did not find significant differences in the fidelity of either enzyme, even when using optimal buffer conditions, thermal cycling parameters, and number of cycles (0.2% and 0.13% error rates for ULTMA and Taq, respectively, after reading about 3,000 bases each). We conclude that for sequencing purposes there is no difference in using a DNA polymerase that contains an inherent 3' to 5' exonuclease activity for DNA amplification. Perhaps the specificity and fidelity of PCR are complex issues influenced by the nature of the target sequence, as well as by each PCR component.
Palavra-chave polymerase chain reaction fidelity
Thermus aquaticus (Taq) DNA polymerase
Thermotoga maritima DNA polymerase
Idioma Inglês
Data de publicação 1998-10-01
Publicado em Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 31, n. 10, p. 1239-1242, 1998.
ISSN 0100-879X (Sherpa/Romeo, fator de impacto)
Publicador Associação Brasileira de Divulgação Científica
Extensão 1239-1242
Fonte http://dx.doi.org/10.1590/S0100-879X1998001000001
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000076614900001
SciELO S0100-879X1998001000001 (estatísticas na SciELO)
Endereço permanente http://repositorio.unifesp.br/handle/11600/679

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