Unit Definition

Taq DNA Polymerase 1.1x Master Mix One unit is defined as the amount of polymerase that
2 mM MgCl2 final concentration incorporates 10 nmoles of dNTPs into acid-precipitable DNA in
30 minutes at 72 °C under standard assay conditions.

Cat. No.: A130301

Protocol
Size Taq DNA Polymerase 1.1x Master Mix,
Cat. No. This protocol serves as a guideline for primer extensions.
Reactions 2 mM MgCl2
Optimal reaction conditions such as incubation times,
ID No. 5200050 temperatures, and amount of template DNA may vary and must
-
Cap colour Blue
be determined individually.
A130301 100 3 x 1.5 ml
Note:
 Set up reaction mixtures in an area separate from that used
for DNA preparation or product analysis. Work on ice at all
times.

1. Thaw Taq 1.1x Master Mix and primers. It is important to

thaw the solutions completely and mix thoroughly before
use to avoid localized concentrations of salts. Keep all
Key Features components on ice.
 High performance thermostable DNA polymerase 2. Set up each reaction. Table 1 shows the reaction set up for a
 Significantly reduced set up time final volume of 50 L. If desired, the reaction size may be
 Diminished risk of contamination scaled down. Use 18 µl of the Taq 1.1x Master Mix in a final
 Increased reproducibility volume of 20 µl.
 No proofreading – lacks a 3'5' exonuclease activity If your template DNA is very dilute, it might be an advantage
 Ideal for TA cloning – leaves an A overhang to use the Ampliqon Taq 2x Master Mix. (See Additional
Products)

Taq DNA Polymerase 1.1x Master Mix is a ready-to-use 1.1x Table 1. Reaction components (reaction mix and template
reaction mix with the Ampliqon Taq DNA polymerase, the NH4
+ DNA)
buffer system, dNTPs and magnesium chloride present. Each Component Vol./reaction Final concentration
reaction requires 45 µl of the 1.1x Master Mix. Simply add Taq 1.1x Master
45 µl 1x
primers, template and water to a total reaction volume of 50 µl Mix
to successfully carry out primer extensions and other molecular Primer A 1 µl (10 µM) 0.2 µM (0.1 – 1.0 µM)
biology applications. Primer B 1 µl (10 µM) 0.2 µM (0.1 – 1.0 µM)
Taq DNA Polymerase 1.1x Master Mix offers several advantages. PCR-grade H2O X µl -
Set up time is significantly reduced. The chance of contaminating genomic DNA: 50 ng (10 – 500 ng)
component stocks is eliminated. Reduction of reagent handling Template DNA X µl plasmid DNA: 0.5 ng (0.1 – 1 ng)
steps leads to better reproducibility. Standard tests can be set bacterial DNA: 5 ng (1 – 10 ng)
up with the confidence that results will be consistent every time.
TOTAL volume 50 µl -

Composition of the Taq DNA Polymerase 1.1x Master Mix (2 3. Mix the reaction mix thoroughly and dispense appropriate
mM MgCl2 final concentration) volumes into reaction tubes. Mix gently, e.g. by pipetting the
reaction mix up and down a few times.
 Tris-HCl pH 8.5, (NH4)2S04, 2.2 mM MgCl2, 0.11% Tween 20
 0.22 mM of each dNTP 4. Add template DNA to the individual tubes containing the
 0.11 units/µl Ampliqon Taq DNA polymerase master mix.
 Stabilizer
5. Program the thermal cycler according to the manufacturer´s
instructions.
Recommended Storage and Stability For maximum yield and specificity, temperatures and cycling
Long term storage at -20 °C. Product expiry at -20 °C is stated on times should be optimized for each new template target or
the label. primer pair.
Option: Store at +4 °C for up to 6 months.
6. Place the tubes in the thermal cycler and start the reaction.
Quality Control
Taq DNA Polymerase is tested for contaminating activities, with Three-step PCR program
no traces of endonuclease activity, nicking activity or Cycles Duration of cycle Temperature
exonuclease activity. 1 2 – 5 minutes 95 °C
25 - 35 20 – 30 secondsa 95 °C
20 – 40 secondsb 50 – 65 °C
30 secondsc 72 °C
1 5 minutesd 72 °C
a.
Denaturation step: This step is the first regular cycling event and Related Products
consists of heating the reaction to 95 °C for 20 – 30 seconds. It causes
melting of the DNA template by disrupting the hydrogen bonds Taq Polymerase (500 units) * Cat. No.
between complementary bases, yielding single-stranded DNA Taq DNA Polymerase 5 U/µl A110003
molecules.  with 10x Ammonium Buffer A111103
 5x PCR Buffer RED A111803
b.
Annealing step: The reaction temperature is lowered to 50 – 65 °C for
20 – 40 seconds allowing annealing of the primers to the single- Taq DNA Polymerase 5 U/µl, RED A200003
stranded DNA template. Typically, the annealing temperature is about  with 10x Ammonium Buffer A201103
3 – 5 °C below the Tm of the primers used. Taq DNA Polymerase 5 U/µl, glycerol free A100003
c.  with 10x Ammonium Buffer A101103
Extension/elongation step: Taq polymerase has its optimum activity
temperature at 72 °C. At this step the DNA polymerase synthesizes a Hot Start Polymerase (500 units) * Cat. No.
new DNA strand complementary to the DNA template strand. The
TEMPase Hot Start DNA Polymerase, 5 U/µl A220003
extension time depends on the length of the DNA fragment to be
 with 10x Ammonium Buffer A221103
amplified. As a rule of thumb, at its optimum temperature the DNA
 5x PCR Buffer RED A221803
polymerase will polymerize a thousand bases per minute.
d. TEMPase Hot Start DNA Polymerase, glycerol free 5 U/µl A240003
Final elongation: This single step is occasionally performed at a
 with 10x Ammonium Buffer A241103
temperature of 72 °C for 5 minutes after the last PCR cycle to ensure
that any remaining single‐stranded DNA is full e tended. High Fidelity - Proof reading (500 units) ** Cat. No.
AccuPOL DNA Polymerase 2.5 U/µl A210003
 with 10x Ammonium Buffer A211103
*Available in kits including one or two buffers (Ammonium Buffer, Standard Buffer
or Combination Buffer). **AccuPOL only available in kits with Ammonium Buffer.
All kits include extra 25 mM MgCl2.
Buffers for DNA polymerases * Cat. No.
10x Ammonium Buffer, 3 x 1.5 ml A301103
10x Standard Buffer, 3 x 1.5 ml A302103
10x Combination Buffer, 3 x 1.5 ml A303103
5x PCR Buffer RED, 6 x 1,5 ml ** A301810
*Ammonium Buffer, Standard Buffer and Combination Buffer are also available as
Mg2+ free buffers, detergent free buffers and Mg2+ and detergent free buffers.
**For direct gel loading and visualisation.

Taq Master Mixes (500 x 50 µl reactions) * Cat. No.

2x Master Mix, 1.5 mM MgCl2 final concentration A140303
2x Master Mix RED, 1.5 mM MgCl2 final concentration A180303
TEMPase Hot Start Master Mixes (500 x 50 µl reactions) * Cat. No.
2x Master Mix A**, 1.5 mM MgCl2 final concentration A230303
2x Master Mix A**BLUE, 1.5 mM MgCl2 final concentration A290403
*Master mixes available also in 1.1x variants as well as 2 mM MgCl2 variants, **Mix
A is Ammonium Buffer based, also available as Mix C based on Combination Buffer.

Special Master Mixes (500 x 50 µl reactions) Cat. No.

Multiplex 2x Master Mix, 3 mM MgCl2 final concentration A260303
GC TEMPase 2x Master Mix I – for GC-rich templates A331703
GC TEMPase 2x Master Mix II – for GC-rich templates A332703
Real-time PCR Master Mixes (400 x 25 µl reactions) Cat. No.
RealQ Plus 2x Master Mix for probe,
 without ROXTM A313402
 with low ROXTM A314402
 with high ROXTM A315402
RealQ Plus 2x Master Mix Green
 without ROXTM A323402
 with low ROXTM A324402
 with high ROXTM A325402
Ultrapure dNTPs* Cat. No.
dNTP Mix 40 mM (2 x 500 µl): 10 mM each dA, dC, dG, dT A502004
dNTP Set, 100 mM each: 250 µl of each dA, dC, dG and dT A511104
*Other concentrations and Single dNTPs are available.

Loading Buffers and Ladders Cat. No.

5x Loading Buffer Red *, 5 x 1 ml A608104
PCR DNA Ladder **, 100 – 3000 bp, 1 x 0.5 ml A610341
* Also available with Blue, Orange or Cyan. ** Available in different size ranges.

Reagents for in vitro laboratory use only.

Other product sizes, combinations and customized solutions are available. Please
look at www.ampliqon.com or ask for our complete product list for PCR Enzymes.
For customized solutions please contact us.
Made in Denmark

Issued 02/2017

Ampliqon A/S, Stenhuggervej 22, DK-5230 Odense M, Denmark. Phone: +45 70201169 Fax: +45 70201179 info@ampliqon.com www.ampliqon.com