Taq DNA Polymerase 5 U/ul, 10x Ammonium Buffer and 10x Standard Buffer


SKU: AO1141 Research Area: Applications:


Taq DNA Polymerase

   10X Ammonium Buffer

10X Standard Buffer

5 units/µl


Cat. No.
10X Ammonium Buffer
10X Standard Buffer
25 mM
1.5 mL
1.5 mL
1.5 mL
2 x 1.5 mL
2 x 1.5 mL
2 x 1.5 mL
4 x 1.5 mL
4 x 1.5 mL
4 x 1.5 mL
3 x 5.0 mL
3 x 5.0 mL
3 x 5.0 mL
6 x 5.0 mL
6 x 5.0 mL
6 x 5.0 mL

Store at -20°C.   For in-vitro laboratory use only


General Description


AS ONE 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.


AS ONE 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 Buffer


Tris-HCl pH 8.5, (NH4)2S04, 15mM MgCl2, 1% Tween 20®


10X Standard Buffer


Tris-HCl pH 8.5, 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 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


Unopened enzyme is stable at -20°C for 2 years after production date.




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 only. Optimal reaction conditions may vary and must be individually determined.


  • In some applications, more than 1.5mM MgCl2 is needed for the best results. For this reason, 25 mM MgCl2 is included with the kit.


  1. Thaw desired 10X Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.


  1. 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 & template DNA)


Vol./reaction Final Conc.
10X Buffer 5 µL 1X
25 mM MgCl2 0 µL (0-7 µL) 1.5 mM                  (1.5–5 mM)
dNTP mix

(12.5 mM of each)

0.8 µL 0.2 mM of

each dNTP

Primer A 1 µL (0.5-5 µL) 0.2 µM                  (0.1–1.0 µM)
Primer B 1 µL (0.5-5 µL) 0.2 µM                  (0.1–1.0 µM)
Taq DNA Polymerase 0.6 µL              (0.2–1 µL) 3 units (1–5 units)
Template DNA Variable Variable
Distilled Water Variable – – – –
TOTAL volume 50 µL – – – –


Table 2.  MgCl2 concentration in a 50 mL 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 rxn (µL):

0 1 2 3 4 5 6


Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a certain MgCl2 concentration is required.


  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.


  1. Add template DNA to the individual tubes containing the master mix.


  1. 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.


  1. Place the tubes in the thermal cycler and start the reaction.



Three- step PCR Program:


Cycles Duration of Cycles Temperature
1 2-5 minutesa 95°C
25-35 20-30 secondsb

20-40 secondsc

30 secondsd




1 5 minutese 72°C









a  Initial denaturation step. Optional, but recommended for genomic DNA

b Denaturation step: Heating the DNA template disrupts the hydrogen bonds yielding ssDNA.

c Annealing step: Primers anneal to ssDNA template. Typically annealing temperature is 3-5°C below the Tm of the primers.

d Extension/elongation step: Taq DNA polymerase synthesizes a new DNA strand complementary to the DNA template. Extension time depends on length of DNA fragment to be amplified. At optimum temperature the DNA polymerase will polymerize 1000 bases per minute.

e Final elongation step: After the last PCR cycle to ensure that any remaining ssDNA is fully extended.


Title Type Size
AO11410x_TaqAmmStd_082415 application/pdf 24 KB

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