Taq DNA Polymerase
Code  BN0130-0500
Classification  PCR試劑 & 1-Step Kit
Size  500u
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PRODUCT DETAIL



Taq DNA Polymerase

With 10X Standard Buffer (MgCl2 15 mM)
With 10X Ammonium Buffer (MgCl2 15 mM)
Conc.: 5 units/μl
Cat. No.: 110104 (1000 Units)
Cat. No.
Size Units
10X Ammonium Buffer
(MgCl2 15mM)
10X Standard Buffer
(MgCl2 15mM)
MgCl2
25 mM
110103
500
1.5 mL
1.5 mL
1.5 mL
110104
1000
2x 1.5 mL
2x 1.5 mL
2x 1.5 mL
110108
10000
6x 5.0 mL
6x 5.0 mL
6x 5.0 mL
Store at -20°C. Reagent for in-vitro laboratory use only
 
General Description
 

Ampliqon Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. The enzyme is isolated from Thermus aquaticus and has a molecular weight of approximately 94 kDa.
Ampliqon Taq DNA Polymerase has both a 5'→3' DNA polymerase and a 5'→3' exonuclease activity. The enzyme lacks a 3'→5' exonuclease activity (no proofreading ability). Taq DNA Polymerase leaves an A′ overhang, which makes the enzyme ideal for TA cloning.
   
10X Ammonium Reaction Buffer
  Tris-HCl pH 8.5, (NH4)2S04, 15 mM MgCl2, 1% Tween 20®.
   
10X Standard Reaction Buffer
  100mM Tris-HCl pH 8.3, 500mM KCl, 15mM MgCl2, 1% Triton X-100.
   
Unit Definition
 
One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.
   
Storage and Dilution Buffer
 
Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20®, 0.5% NP40, 50% glycerol.
   
Quality Control
 
Each lot of Taq DNA Polymerase is tested for contaminating activities, with no trace of endonuclease activity, nicking activity, exonuclease activity or priming activity.
 
Suggested Protocol using Taq DNA Polymerase
 
This protocol serves as a guideline for primer extensions. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.
   
  Notes:
 
  • Set up reaction mixtures in an area separate from that used for DNA preparation or product analysis.
  • 15 mM MgCl2 is present in the 10X buffer. The 1X concentration is 1.5 mM MgCl2.
  • In some applications, more than 1.5 mM MgCl2 is needed for the best results. For this reason, 25 mM MgCl2 is included with the kit. Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.
   
  1. Thaw 10X Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.
  2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.
   
  Table 1. Reaction components (master mix and template DNA)
 
Component Vol./reaction Final Conc.
10X Buffer 5 μL 1X
dNTP mix
(12.5 mM of each)		 0.8 μL 0.2 mM of each dNTP
Primer A Variable 0.1–1.0 μM
Primer B Variable 0.1–1.0 μM
Taq DNA Polymerase Variable 1-5 units
Distilled Water Variable - - - -
Template DNA Variable Variable
TOTAL volume 50 μL - - - -
   
  Table 2. MgCl2 concentration in a 50 μL reaction
 
Final MgCl2 conc. in reaction (mM)
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Additional volume of 25 mM MgCl2
per reaction (μL):
0
1
2
3
4
5
6
   
  1. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently, e.g., by pipetting the master mix up and down a few times.
  2. Add template DNA to the individual tubes containing the master mix.
  3. Program the thermal cycler according to the manufacturer’s instructions.
    For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.
  4. Place the tubes in the thermal cycler and start the reaction.
   
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  Tween 20® is a registered trademark of ICI Americas, Inc.
   
  NOTICE
 
In certain countries, patents cover the PCR process. This product is intended for researchers having a license to perform PCR or those not required to obtain a license.