SPEED Hi-Fidelity Taq DNA Polymerase PCR Kit

SPEED Hi-Fidelity Taq DNA Polymerase PCR Kit

SKU Description Size Price Shipping  
LYM1008 SPEED Hi-Fidelity Taq DNA Polymerase PCR Kit 500 U $180 2-5 Days Add To Shopping Cart
Datasheet:
The thermostable enzyme replicates DNA at 75ºC. It catalyzes polymerization of nucleotides into duplex DNA in the 5'-->3' direction in the presence of magnesium. The enzyme exhibits 3'-->5' exonuclease (proofreading) activity and is recommended for use in PCR and primer extension reactions that require high fidelity. Amplification with this polymerase generates blunt-ended PCR products.

Storage

Keep at -20°C for long term storage. 

Enzyme Storage Buffer

50mM Tris-HCl (pH 8.2 at 25°C), 0.1mM EDTA, 1mM DTT, 0.05% CHAPS and 50% glycerol.

10X PCR Buffer

200mM Tris-HCl (pH 8.8), 100mM KCl, 100mM (NH4)2SO4, 1% TritonX-100 and enzyme stablizer. 25mM MgCl2 supplied separately.


Protocol

(This is a general guide. Specific conditions should be optimized by the end users)

1. Add the following components to a sterile tube siting on ice

           Components                                         Volume

          DNA Template                                        ~100ng
          Forward Primer (10 µM)                           1µl
          Reverse Primer (10 µM)                           1µl
         10X PCR Buffer                                         5µl
         25mM MgCl2                                             3µl
         dNTP mixture (10mM each)                      1µl
         Enzyme                                                    0.5-1µl
          H2O                                                      to 50µl


2. Mix contents of tube and centrifuge brie?y.

3. Incubate tube in a thermal cycler at 94°C for 3 min to completely denature the template.

4. Perform 30-40 cycles of PCR ampli?cation as follow:

             Denature: 94°C for 30 sec
             Anneal:    55°C for 30 sec (annealing temperature may need to be adjusted based on Tm)
             Extend:    72°C for 1 min/1kb template

5. Analyze the ampli?cation products
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