A strong strand displacement activity of thermostable DNA polymerase markedly improves the results of DNA amplification

Biotechniques. 2014 Aug 1;57(2):81-7. doi: 10.2144/000114198. eCollection 2014 Aug.

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.

Keywords: DNA polymerase; LAMP; PCDR; PCR; isothermal amplification; quantitative amplification; real-time amplification; strand displacement.

MeSH terms

  • Animals
  • DNA / genetics
  • DNA / isolation & purification*
  • DNA Primers / genetics
  • Mice
  • Mycobacterium tuberculosis / genetics
  • Nucleic Acid Amplification Techniques / methods*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Taq Polymerase / chemistry
  • Taq Polymerase / genetics*

Substances

  • DNA Primers
  • DNA
  • Taq Polymerase