Expansion of repertoire of modified DNAs prepared by PCR using KOD Dash DNA polymerase

Org Biomol Chem. 2005 Jul 7;3(13):2463-8. doi: 10.1039/b504330a. Epub 2005 Jun 9.


Thymidine analogues bearing a variety of functional groups at the C5-position via an amino-linker arm were prepared and the substrate activity for PCR using thermophilic KOD Dash DNA polymerase was examined. The enzyme accepted the thymidine analogues bearing pyridine, imidazole, biotin, a cationic-charged guanidinium, a cationic-charged amino, mercaptopyridyl and phenanthrolne groups at the C5-position, forming the corresponding PCR product. However, a thymidine analogue bearing a carboxyl group at the C5-position was a poor substrate and the corresponding PCR products could not be obtained. The thymidine analogue bearing a mercapto group was also a poor substrate for the enzyme, because it dimerized by disulfide linkage under PCR conditions. The enzyme hardly accepts the thymidine analogues with a negatively-charged carboxyl group or a bulky group as a substrate. KOD Dash DNA polymerase, having a broader substrate specificity than any other DNA polymerase, will expand the variety of modified DNAs that can be prepared by PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amines / chemistry
  • Biotin / chemistry
  • DNA / chemical synthesis*
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism*
  • Guanidine / chemistry
  • Imidazoles / chemistry
  • Polymerase Chain Reaction
  • Pyridines / chemistry
  • Substrate Specificity
  • Thymidine / analogs & derivatives
  • Thymidine / chemistry*


  • Amines
  • Imidazoles
  • Pyridines
  • Biotin
  • imidazole
  • DNA
  • DNA-Directed DNA Polymerase
  • Guanidine
  • pyridine
  • Thymidine