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Taq DNA Polymerase

 
Description: Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity and a double-strand specific 5´→3´ exonuclease activity. It is supplied with 10x Standard Taq Reaction Buffer and MgCl2, which is detergent-free and designed to be compatible with existing assay systems.
 
Source: Recombinant Taq DNA polymerase purified from an E. coli strain carrying Thermus aquaticus gene.
 
Applications
  • PCR
  • Primer Extension
  • Microarray Analysis
  • High-Throughput PCR
 
Enzyme Properties
Heat Inactivation: No

Reaction Conditions:
1x Standard Taq Reaction Buffer:
     10 mM Tris-HCl
     50 mM KCl
     1.5 mM MgCl2
     pH 8.3 @ 25°C

Unit Definition: One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.

Concentration: 5,000 units/ml
 
Storage Temperature: -20°C
 
Protocol
  1. Gently mix the reaction and spin down in microcentrifuge. If the thermocycler does not have a heated cover, add one drop of mineral oil to the reaction tube to prevent evaporation.
 
Component
25 µl
50 µl
Final Conc.
Reaction Buffer (10x)
2.5 µl
5 µl
1x
MgCl2 (30 mM) or
MgSO4 (20 mM)
1.25 µl
1.5  µl
2.5 µl
3 µl
1.5 mM
dNTPs Mix (10 mM, 2.5 mM/each)
0.5 µl
1 µl
200 µM
5’ Primer (10 µM)
0.5 µl
1 µl
0.2 µM
3’ Primer (10 µM)
0.5 µl
1 µl
0.2 µM
DNA template
Determined by user
0.1–1 ng/ml plasmid DNA
1-10 ng/ml genomic DNA
Taq
1-1.5 U
2-3 U
 
Nuclease free H2O
to 25µl
to 50 µl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  1. Cycling conditions for a routine PCR Reaction:
 
Initial Denaturation
94-95oC
2-5 min
25-40 cycles
94-95oC
45-70oC
72oC
15-30 sec
15-30 sec
1 min per 1000 base pairs
Final Extension
72oC
5 min
Final Soak
4oC

 
 
 
 
 
 
 
 
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