Hot-Start PCR: M-Superhot Taq Polymerase

Superior Taq Polymerase for Real Time PCR and Hot-Start PCR, low-copy number PCR, PCR of difficult templates, Hot-Start activity (only 5 min initial denaturation). The enzyme is developed to enhance the specificity, sensitivity and yield of DNA amplification. The enzyme provides a convenient setting up at room temperature

Cat.-no Description Amount Price € Shop
 S105  Maximo M-Superhot Taq DNA Pol. (qPCR)     200 units     69.00
   55.00
 add 
 S106  Maximo M-Superhot Taq DNA Pol. (qPCR)  5x200 units  279.00
 239.00
 add 
 S107  Maximo M-Superhot Taq DNA Pol. (qPCR)    5000 units  995.00
 895.00
 add

m-Superhot Taq DNA Polymerase for real-time PCR and Hot-Start PCR: Order/request by E-mail
m-Superhot Taq DNA Polymerase for real time PCR and Hot-start PCR: Datasheet
m-Superhot Taq DNA Polymerase for real-time PCR and Hot start PCR: Deutsche Beschreibung

Features: Maximo M-Superhot Taq DNA Polymerase for qPCR is designed for Real-Time PCR and Hot-start PCR. A special inhibitor suppresses the reaction at room temperature until after the first denaturation step. This prevents primer-dimers and other artefacts. Using the enzyme there is no need to adjust the existing standard PCR protocol.

Applications: 
- Hot Start and real time PCR
- Multiplex PCR
- Amplification of complex genomic and cDNA templates
- no primer-dimers and other artêfacts; inactive at room temperature
- short activation time for real time PCR
- enhanced PCR sensitivity

Description:
Maximo M-Superhot Taq DNA for qPCR and Hot-Start-PCR is an optimized mixture of a high processive Taq DNA Polymerase and special inhibitors to Taq DNA for real time PCR. The enzyme is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity and a double-stranded specific 5´→3´ exonuclease activity. The enzyme consists of a single polypeptide with a molecular weight of 94kDa. It is developed for real time PCR or as basis enzyme for  real time PCR diagnostics systems.
 
Concentration: 5 u/µl

Unit definition:
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP's into acid-insoluble form in 30 minutes at 74oC under assay conditions:
25mM TAPS pH 9.3 at 25oC, 50mM KCl, 2mM MgCl2; 1mM beta-mercaptoethanol; 200µM each dATP, dGTP, dTTP and 100 µM dCTP (a mix of unlabled and µ-[32P]-labled); 12.5 µg activated salmon sperm DNA in the final volume of 50 µl

Storage Buffer:
20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA; 1 mM DTT, 50 % Glycerol, 0.5 % Nonident P-40, 0.5 % Tween-20

Reaction Buffer:
Reaction buffer (10X)” incomplete” (red cap):160 mM (NH4)2SO4, 670mM TrisHCl pH8,8, 0,1% Tween-20
Reaction buffer (10X) “complete” (yellow cap):160 mM (NH4)2SO4, 670mM TrisHCl pH8,8, 0,1% Tween-20, 25mM MgCl2
separate Tube: MgCl2 (100 mM, green cap)

Quality control:
Activity and performance test in real time PCR, SDS-PAGE purity, absence of endonucleases/nickases and exonucleases test
 
Usage:  

Use your existing and optimized protocol. In contrast to chemically modified Taq DNA pol. where the first denaturation step needs up to 15 min, Maximo M-Superhot Taq for Real Time PCR does not need a prolonged heating or denaturation time and works excellent basis enzyme for real time PCR.

Components Volume per reaction
 10X reaction buffer  10 µl
 100 mM MgCl2  optional
 dNTP-Mix (40mM)  1.0 µl
 Up-stream primer (10 µM stock)  0,5-2.5 µl
 Down-stream primer (10µM stock)   0.5-2,5 µl
 Template DNA  0.1-15 ng/ml plasmid DNA
 1-10 µg/ml genomic DNA
 Maximo M-Superhot Taq DNA (5 u/µl)   0.2 - 1.0 µl  
 Sterile dest. Water (molecular grade)  up to 50 µl total reaction volume

Note: 
- vortex all solutions carefully before using
- add the enzyme after Template DNA
- may you have to optimize the MgCl2 concentration for best result

General Thermo-Cycler protocol:
 Step  Time  Temperature
 Initial denaturation  2-5 min  94-95°C
 
25-30 Cycles:
 Denaturation
 Annealing
 Extension
 
 10-25 sec
 10-25 sec
 60 sec

 94-95°C
 55-65°C
 72°C per 1kb
 
 Final extension    5 min  72°C

Note: 
- In case of low amount of DNA template, additionally cycles may be used 

Transportation: on blue ice

Storage: at -20°C for 24 months or  for more than 3 months at +4°C


Related products for Hot start PCR:
p-Superhot Taq Polymerase (hot-start PCR)

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