Furthermore, we hope that this report will remind laboratorians of the need to understand the amplification characteristics of the thermostable DNA polymerases
that they use for PCR-based assays and to periodically review frequently updated SNP databases for uncommon polymorphisms that could disrupt amplification of any given target sequence.
It also inhibits B-family DNA polymerases
from eukaryotes [18-21], bacteria  and viruses [14; 23-25].
KlenTaq 1 (DNA Polymerase
Technology) and Taq (New England BioLabs) were used in this study.
Inorganic pyrophosphatase (40 U/ml) and Sequenase version 2.0 T7 DnA polymerase
(13 U/ml) were obtained from USB Corporation (Cleveland, OH, USA).
As an enzyme for PCR, we used the following types of DNA-dependent DNA polymerases
: Taq DNA polymerase
, Tersus DNA polymerase
, Deep Vent DNA polymerase
and Phusion DNA polymerase
Hogrefe, "PCR fidelity of Pfu DNA polymerase
and other thermostable DNA polymerases
," Nucleic Acids Research, vol.
The concentrations used for each DNA polymerase
are specified in the respective figure legends.
However, researchers have been trying to produce such enzyme from various versions of Taq DNA polymerase
gene via genetic engineering.
Another tenable explanation for the presence of multiple specialised DNA polymerases
in vertebrates is that each evolved to accurately bypass a particular type of naturally occurring base damage.
Thus, DNA polymerases
might sensitively recognize the [gamma]-amidotriphosphate moiety of the substrates, and the geometrical fluctuation caused by non-cognate pairings, such as Ds-Ds and A-Pa, significantly reduces the interaction between the amino acid residues in the polymerase and the [gamma]-amidotriphosphate moiety, resulting in the loss of the function of the amidodiphosphate part as a leaving group, like the pyrophosphate moiety of triphosphate substrates.
Cloning and sequencing of lcr1 by Pfu DNA polymerase
: To determine whether the sequence variants were created during PCR due to Taq polymerase error, lcr1 was amplified from the parasite genomic DNA by Pfu DNA polymerase
(Fermentas Co., Ontario, Canada) which exhibits 3'[right arrow]5' exonuclease (proof reading) activity according to manufacturer's instructions.
Studies will determine the in vivo roles of DNA polymerases
and other proteins likely to be involved in UV mutagenesis.