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A strong strand displacement activity of thermostable DNA polymerase markedly improves the results of DNA amplification.
Ignatov, Konstantin B; Barsova, Ekaterina V; Fradkov, Arkady F; Blagodatskikh, Konstantin A; Kramarova, Tatiana V; Kramarov, Vladimir M.
Afiliação
  • Ignatov KB; Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; All-Russia Institute of Agricultural Biotechnology, Moscow, Russia.
  • Barsova EV; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Evrogen JSC, Moscow, Russia.
  • Fradkov AF; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Evrogen JSC, Moscow, Russia.
  • Blagodatskikh KA; All-Russia Institute of Agricultural Biotechnology, Moscow, Russia; Syntol JSC, Moscow, Russia.
  • Kramarova TV; The Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden.
  • Kramarov VM; Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; All-Russia Institute of Agricultural Biotechnology, Moscow, Russia.
Biotechniques ; 57(2): 81-7, 2014 Aug.
Article em En | MEDLINE | ID: mdl-25109293
ABSTRACT
The sensitivity and robustness of various DNA detection and amplification techniques are to a large extent determined by the properties of the DNA polymerase used. We have compared the performance of conventional Taq and Bst DNA polymerases to a novel Taq DNA polymerase mutant (SD DNA polymerase), which has a strong strand displacement activity, in PCR (including amplification of GC-rich and complex secondary structure templates), long-range PCR (LR PCR), loop-mediated amplification (LAMP), and polymerase chain displacement reaction (PCDR). Our results demonstrate that the strand displacement activity of SD DNA polymerase, in combination with the robust polymerase activity, provides a notable improvement in the sensitivity and efficiency of all these methods.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Reação em Cadeia da Polimerase / Taq Polimerase / Técnicas de Amplificação de Ácido Nucleico Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Biotechniques Ano de publicação: 2014 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Reação em Cadeia da Polimerase / Taq Polimerase / Técnicas de Amplificação de Ácido Nucleico Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Biotechniques Ano de publicação: 2014 Tipo de documento: Article