title: A strong strand displacement activity of thermostable DNA polymerase markedly improves the results of DNA amplification.
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.
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; All-Russia Institute of Agricultural Biotechnology, Moscow, Russia.
authors.forename: Konstantin B
authors.initials: KB
authors.lastname: Ignatov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Evrogen JSC, Moscow, Russia.
authors.forename: Ekaterina V
authors.initials: EV
authors.lastname: Barsova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Evrogen JSC, Moscow, Russia.
authors.forename: Arkady F
authors.initials: AF
authors.lastname: Fradkov
authors.affiliation: All-Russia Institute of Agricultural Biotechnology, Moscow, Russia; Syntol JSC, Moscow, Russia.
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Blagodatskikh
authors.affiliation: The Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden.
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Kramarova
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; All-Russia Institute of Agricultural Biotechnology, Moscow, Russia.
authors.forename: Vladimir M
authors.initials: VM
authors.lastname: Kramarov
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000737:chemistry / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004247:DNA; D019914:Taq Polymerase
keywords: DNA polymerase; LAMP; PCDR; PCR; isothermal amplification; quantitative amplification; real-time amplification; strand displacement