DFS-Taq "Hot" is the " Hot Start PCR " version of DFS Taq DNA Polymerase that provides a new formula in buffers and additives to prevent failures in PCR-applications were inhibitors (e.g.
proteins, fat or PS) reduce the performance.
The Taq Polymerase is blocked with monoclonal Antibodies. Both components, the Taq Polymerase and the Antibody is available, separatly at GeneON's shop.
The robust hot start polymerase is well suited for sensitive experiments using random primers or bacterial templates. Because of the high sensitivity less than 6 molecules can be detected.
- reliable and reproducable quantification in qPCR
- perfect for real time PCR
- especially for diagnostic purposes
- reaction set-up at room temperature
- activation of enzyme during first heating
- no change or optimization of protocol necessary
- high specifity, reduced primer mismatch or dimers
Instead of conventionally purified Taq-DNA Polymerase for sensitive PCR reactions, for the detection of bacterial DNA or for applications where inhibitors decrease the performance of regular
- Hot start PCR
- Real time PCR
- Amplification of complex genomic and cDNA templates
- Multiplex PCR
- High specifity PCR
Same as for conventionally purified Taq-DNA Polymerase.
5 units/μl supplied in 10 mM KPO4 (pH 7.4 at 25°C), 0.1 mM EDTA, 0.1% Tween 20, 0.1% Triton-X 100 and 50 % (v/v) glycerol.
One unit is defined as the amount of enzyme required to incorporate 10 nmoles of dNTP into acid-insoluble material in 30 min at 74°C.
Reaction Buffers provided:
Ammonium-Reaction buffer (10X)” incomplete”
Ammonium-Reaction buffer (10X) “complete” with 25mM MgCl2
MgCl2 (100 mM)
Transport: with "blue ice" or at ambient temperature.
Storage: at –20 °C is recommended to safeguard against growth of bacteria that may be introduced during handling.
- Endonucleases Incubation of 20 units of the enzyme in 1x reaction buffer
with 1 μg lambda DNA for 16 h
at 65°C in 50 μl yields no detectable degradation of DNA.
- Incubation of 20 units of the enzyme in 1x reaction buffer with 1 μg
lambda DNA EcoR I/Hind III fragments
for 16 h at 65°C in 50 μl yields no detectable degradation of DNA.
- Incubation of 32 units of the enzyme in 1x reaction buffer with 1 μg
supercoiled pUC18 DNA for 16 h at 70° C in 50 μl resulted in no
- Priming activity Incubation of 40 units of the enzyme in 1x reaction buffer
with 100 ng template DNA and
0.2 mM dNTPs each, but without primers in 100 μl resulted in no DNA
- PCR Test Good performance of DNA amplification was confirmed by
using Lambda DNA as template (amplified fragment 12 kb) and human
placenta DNA as template (amplified fragment 3.0 kb).
- No DNA contamination with enterobacterial DNA