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SuperTaq

Product: SuperTaq DNA Polymerase
Item Number: Super Taq
Conc:5units/µl

Product Information:
SuperTaq is relatively more thermostable DNA Polymerase isolated from a strain of Thermus aquaticus. It has a half life of 3 hours at 95 °C, it is very stable. It has high fidelity with an error frequency 10 / 106 (or 0.01 / 103) during DNA synthesis. SuperTaq is designed for use in primer extension reaction. DNA sequencing at high temperature may decrease the second structure of some DNA templates and permit polymerization through base-paired region. DNA sequencing with Taq DNA Polyinerase produces uniform bands intensities and low background.

Quality& Performance Parameters:
SuperTaq is highly purified free of contaminating endonucleases, exonucleases and nicking activity. For endonuclease assay, 1 g of Lambda I Hind III DNA is incubated with 20 units of the enzyme in assay buffer at 75º C for 16 hrs and no visible contaminating activity is observed; For exonucleases assay, 1 g of pBR322 plasmid DNA is incubated with 10 units of enzyme for 16 hrs at 75º C in assay buffer and no detectable exonuclease is observed. The purity of the enzyme is also evaluated by adding 10 units of SuperTaq in 100 l of a reaction mixture for making first strand cDNA at beginning and no impaired effect on the first strand is observed.
Unit :
One unit incorporates 10 nmole of dNTP into acid-insoluble material in 30 mm. at 74º C.
Concentration in Storage Buffer: 5 units / p1 in 100mM KC1, 20 mM Tris HC1 (pH 8.0, 22°C ), 0.lmM EDTA. 0.5mM PMSF, 1mM DTT, 50% glycerol.
10 X Taq Reaction Buffer:
(New!) 100 mM KC1, 100mM (NH4)2SO4, 200 mM Tris HC1 (pH 8.75) at 22° C, 1% Triton X-100 and 1mg/ml BSA. Buffer is optimized for use with 200 M dNTPs.
Magnesium Sulfate:
(New!) 20mM MgSO4. The final MgSO4 may be variable according to requirements. Normally 2mM MgSO4 is recommended.
General Reaction mixture for PCR:
Taq (5u/ul): 0.5ul, l0xRxn Buffer: l0ul, MgSO4 (20mM): l0ul, dNTP mixture (2.5mM each): 8u1, Primer 1: 0.2-1.0um, Primer 2 : 0.2-1 .0uM, Template : 10pg-lug, Sterilized ddH2O up to l00ul
Primer Extension Characteristics:
Taq has the independent terminal transferase activity which results in the addition of a single nucleotide ( adenosine ) at 3' end of the extension product. TA cloning vector is recommended if the extension product is needed to be cloned.

Reaction Mixture -Set Up
  • Gently vortex and briefly centrifuge all solutions after thawing.
  • Add components, in the following order, into a thin-walled PCR tube. Keep all components on ice. The following control PCR reactions should be performed in parallel with your experiments to ensure that the Super Taq is working properly.

Reagent Final Concentration Quantity
Water (PCR--Grade) --- Variable
10x Taq reaction buffer 1x 5µl
MgSO4(20mM) 2-4mM variable
2.5mM dNTP mixture 200µM of each 4µ1
Primer I, forward 0.l-1µM variable
Primer II, reverse 0.1-1µM variable
Super Taq 1-1.5u/50µl variable
Template DNA See note 1 variable
Total Volume --- 50µl
 
Reagent Positive Control Negative Control
Water (PCR--Grade) 32.8µl 33. 8µl
10x Taq reaction buffer 5µl 5µl
MgSO4(20mM) 5µl 5µl
2.5mM dNTP mixture 4µl 4µl
Primer I, forward lµl lµl
Primer II, reverse 1 µ1 1 µ1
Super Taq 0.2µ1 0.2µ1
Template DNA 1µ1 ---
Total Volume 50µl 50µl
 
 

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