|
FEATURES
-
Lower
error rates than Taq (Taq error rate 1/6 Kbp, Tth error rate
1/10 Kbp)
-
The
same enzyme performs the two steps in a RT-PCR reaction
-
Reverse
transcription activity is optimal at high temperatures (over
50 ºC and up to 70 ºC), therefore
avoiding secondary structures present in RNA (in contrast to
viral reverse transcriptases, such as AMV and M-MLV, where low
reaction temperatures do not denature secondary structures)
-
Half
life at 94 ºC is 40 min. Polymerase activity remains
unaltered after more than 40 cycles of amplification
-
Radioactively
labelled dNTPs, as well
as biotin, fluorescein and digoxigenin-labelled dNTPs (including
dUTP) can be used as substrates
-
Reaction
products can be directly used in T/A cloning
Applications
-
Routine
RT-PCR up to 1 Kbp
-
RT-PCRs
where RNA may have secondary structures and affect reaction
yield
-
RT-PCRs
with limiting amounts of template
-
Cyclic
protocols and Enhancing Buffer included for maximum yield
Description
-
Modified,
recombinant form of the DNA Polymerase from Thermus
thermophilus HB27, expressed in Escherichia
coli, and purified by non-chromatographic methods
-
The
purified enzyme possesses reverse transcriptase activity, as
well as 5’-3’ polymerase activity, as well as weak
5’-3’ exonuclease activity. No 3’-5’ activity is
detected (it is not a proof-reading enzyme)
-
Optimised
for reverse transcription, while retaining the polymerase
activity. No RNase H activity is associated with the reverse
transcriptase activity
-
Performance
of the whole process in one single tube dramatically lowers
risk of contaminations, avoids secondary structures, and total
reaction time is around 2 hours
|
Standard
buffer |
Mg
free buffer |
- |
|
S53941 |
S53981 |
25
rxn |
|
S53942 |
S53982 |
50
rxn |
|
S53943 |
S53983 |
100
rxn |
|
Standard
buffer – gel form* |
Mg
free buffer – gel form |
- |
|
S54091 |
S54111 |
25
rxn |
|
S54092 |
S54112 |
50
rxn |
|
*
DNA polymerase in gel form has the same properties as its liquid
counterpart, but it also presents the following advantages:
-
Allows
shipment at room temperature, avoiding problems due to delay
-
Allows
storage at room temperature (maximum 3 weeks), at 4 ºC
-
Allows
hot-start
reactions without using special enzymes
-
Customised
packaging:
Polymerase
in one vial: resuspend with included buffer for use Pre-aliquoted
reactions: catalog references include polymerase, buffer
and dNTPs. Only add water, your sample and primers! If you
would like to include also your primers, please do not
hesitate to ask us for a customised reference
|
|
|
