Polymerases
for PCR
The need
for innovative application-specific thermostable DNA polymerases is growing
rapidly. Qbiogene has developed a comprehensive selection of enzymes covering
all major PCR applications. Several of these polymerases are completely
unique and offer important advantages. The tables below describe the major
features and benefits of each enzyme.
 Taq
DNA Polymerase |
For
standard PCR applications – highly reliable, well-established
and widely used polymerase. |
| Q-BioTaq
DNA Polymerase |
Recombinant
truncated form of Taq DNA Polymerase lacking 5’-3’exo activity.
|
| SurePrime
Polymerase |
Chemically
modified Taq DNA Polymerase for convenient "hot start" PCR. |
| Isis
DNA Polymerase™ |
Highly
accurate, high thermostability proofreading recombinant DNA Polymerase
originally isolated from Pyrococcus abyssii. Error rate is approximately
1 per 1.6 million base pairs per PCR cycle. |
| Arrow
Taq™ DNA Polymerase |
High
performance mix for high sensitivity and long template amplification.
Accuracy sufficient for most cloning needs. |
| |
Taq
DNA
polymerase |
SurePrime™
|
Isis
DNA polymerase |
Arrow™
|
Q-bioTaq™
|
Feature/
characteristic |
General
PCR |
Hot Start |
High Fidelity
|
High Sensitivity
/ Long fragment PCR |
Multiplex PCR
and RAPDs |
| 5' -›
3' exonuclease |
|
|
- |
|
- |
| 3' -›
5' exonuclease |
- |
- |
|
|
- |
| Error
rate (106) |
24 |
24 |
0.6 |
18 |
24 |
Thermostability
(half life at 95°C) |
40 min |
40 min |
18h |
40 min |
80
min |
| Residual
polymerase activity at 25°C |
yes |
no |
yes |
yes |
yes |
| Longest
amplicons |
Up to
7 kb |
Up to
7 kb |
Up to
10 kb |
Up to
21 kb |
Up
to 7 kb |
| 3' -
end |
dA |
dA |
blunt |
dA/blunt
mix |
dA |
Quality
Control
To ensure the highest standard of quality, each lot of thermostable DNA
polymerase is checked for activity, function and purity. Each thermostable
polymerase is shipped with a lot-specific quality control datasheet.
Absence
of nickases
Confirmed by incubating increasing amounts of enzyme with supercoiled
plasmid DNA (pBR322). The maximum number of units that results in no relaxation
of the supercoiled DNA, as visualized on an agarose gel, is stated on
the lot-specific data sheet.
Absence
of endonuclease contamination
Verified by an assay with Hind III/EcoR I fragments of DNA. Increasing
amounts of enzyme are incubated with constant amount of substrate under
appropriate incubation conditions. The maximum number of units that results
in no degradation of the fragment pattern on agarose gel electrophoresis
is stated on the lot-specific data sheet.
Absence
of 3' exonuclease activity
Confirmed on 32P Klenow labeled Hind III fragments of DNA. After incubation
under appropriate conditions, the absence of released radioactive phosphate
is checked by means of DE-81 adsorption.
Absence of 5' exonuclease and 5' phosphatase activity
Demonstrated
on 32P polynucleotide kinased Hind III fragments of DNA. After incubation
under appropriate conditions, the absence of released radioactive phosphate
is checked by means of DE-81 adsorption.
Absence
of ribonucleases
Checked on 32P labelled RNA. After incubation under appropriate conditions,
the absence of released 32P is checked using DE-81 adsorption.
PCR assay
1
Carried out using human genomic DNA as a template (ß-globin gene)
with decreasing amounts of template DNA and decreasing units of DNA polymerase.
A specific PCR product of 420 bp must be obtained.
PCR assay
2
Carried out using phage DNA as a template ( DNA) with decreasing amounts
of template DNA and decreasing units of DNA polymerase. A specific PCR
product of 500 bp must be obtained.
Purity
Verified by SDS-PAGE.
Patent
Information for PCR Process
Patent
Information for PCR Process
|
