Taq Polymerase 

Thermostable DNA Polymerase

Thermus aquaticus, recombinant, E. coli

CATALOG# PCR-211

For general laboratory use.

Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles 

of dNTP's into an acid-insoluble form in 30 minutes at 70 °C using hering sperm DNA as substrate.

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C

avoid freeze/thaw cycles

Shelf Life: 12 months

Form: liquid

Concentration: 5 units/μl

Description:

Taq Polymerase is recommended for routine PCR applications (up to 4 kb fragment length), high throughput PCR or 

genotyping. The buffer system guarantees robust and reliable amplification results in almost all PCR applications. 

The Crystal Buffer contains a well-balanced ratio of potassium-, ammonium- and magnesium-ions to ensure high 

specificity and minimal by-product formation without the need of additional optimization steps.

Ruby Buffer additionally contains gel loading buffer and an inherent red dye. The red dye allows an easy visual 

control during PCR set-up and in combination with the density reagent the direct loading of the reaction product into the gel.

The enzyme replicates DNA at 72 °C. It catalyzes the polymerization of nucleotides into duplex DNA in 5'→3'

 direction in the presence of magnesium. It also possesses a 5'→3' polymerization-dependent exonuclease replacement 

activity but lacks a 3'→5' exonuclease (proof-reading) activity.

Content:

Taq Polymerase (red cap)

5 units/μl Taq DNA Polymerase in 20 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5 % Tween-20, 0.5 % 

Nonidet P-40, 50 % (v/v) Glycerol, pH 8.0 (25°C)

Ruby Buffer (black cap)

10 x conc. complete PCR buffer containing 200 mM Tris-HCl, KCl, (NH4)2SO4 and 20 mM MgCl2, red tracking 

dye and density reagent for gel loading

Crystal Buffer (green cap)

10 x conc. complete PCR buffer containing 100 mM Tris-HCl, KCl, (NH4)2SO4 and 15 mM MgCl2

component PCR-211S PCR-211L         PCR-211XL

Taq

Polymerase 200 units/ 40 μl 1000 units/ 5 x 40 μl 10000 units/ 2 x 1 ml

Ruby Buffer 1.2 ml 5 x 1.2 ml          50 ml

CrystalBuffer 1.2 ml 5 x 1.2 ml          50 ml

Assay Set-Up:

Before starting, vortex all components thoroughly to ensure homogeneity.

Prepare a premix for the number of assays you need according to the following protocol:

comp.           cap stock conc. final conc.              1 assay @ 20 μl 1 assay @ 50 μl

PCR-grade Water           white                                               fill up to 20 μl fill up to 50 μl

Ruby Buffer or Crystal Buffer    black or green 10x          1x                       2 μl          5 μl

dNTP Mix / 10 mM #NU-1006    white 10 mM  200 μM                       0.4 μl          1 μl

Taq Polymerase            red 5 units/μl          0.025 units/μl               0.1 μl          0.25 μl

primer mix or each primer        10 μM each primer  200 - 400 nMeach primer     0.4-0.8 μl                  1 - 2 μl

template

/sample DNA                                               10 μl < 10 ng DNA 20 μl < 20 ng DNA

Select PCR tubes, stipes or plates as recommended for your cycler model.

Aliquot premix into each well and add template DNA (or PCR-grade Water for negative controls).

Cycling Conditions:

Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.

initial

denaturation 95 °C       2 min        1x

denaturation 95 °C                             10 - 20 sec 25 - 35x

annealing1) 50 - 68 °C               10 - 20 sec

elongation2) 72 °C               20 sec - 4 min

1)The annealing temperature depends on the melting temperature of the primers used.

2)The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.

Gel Loading and Down-Stream Applications:

Ruby Buffer (#PCR-272) includes a density reagent + tracking dye and allows the direct loading of the PCR products into 

a electrophoresis gel. For DNA detection / fluorescent DNA staining we recommend to use new generations dyes 

(i.g. GREEN DNA Stain, #PCR-273) instead of the classical but highly mutagenic ethidium bromide.

Crystal Buffer(#PCR-271) is recommended for down-stream applications such as DNA sequencing, ligation, restriction 

digestion or where an analysis of the PCR product by absorbance or fluorescence excitation is required. For gel 

electrophoresis add gel loading buffer and fluorescent DNA stain (i.g. Gel Loading Buffer with DNA Stain, #PCR-274 - 

#PCR-276) before loading the PCR into the gel. Using pre-stained gels or post-run staining protocols is also possible.

Additional Buffer Systems:

Labeling Buffer (#PCR-263) is recommended for DNA labeling or mutagenesis applications. The buffer is specially 

optimized for incorporation of labeled or modified nucleotides into DNA. It gives superior results in a broad range 

of reaction conditions with most primer-template pairs but amplification may also tend to an increased unspecifity.

KCl Buffer (#PCR-262) is recommended for use in routine PCR reactions. The buffer is optimized for highest specificity 

but may require additional fine-tuning of assay parameters like MgCl2 concentration and annealing temperature.

Optimization of MgCl2 concentration:

A final Mg2+ concentration of 2.0 mM is recommended in combination with Crystal Buffer. However, if an individual

 Mg2+ optimization is essential add 25 mM MgCl2 stock solution (#PCR-266) as shown in the table below.

final MgCl2 conc. 20 μl final assay volume 50 μl final assay volume

1.5 mM - -

2 mM 0.4 μl 1.0 μl

3 mM 1.2 μl 3.0 μl

4 mM 2.0 μl 5.0 μl

5 mM 2.8 μl 7.0 μl

Related products:

dNTP Mix / 10 mM, #NU-1006

Ruby Buffer, #PCR-272

Crystal Buffer, #PCR-271

KCl Buffer, #PCR-262

Green DNA Stain, #PCR-273

Gel Loading Buffer with DNA Stain: Blue #PCR-273, Green #PCR-275, Orange #PCR-276

Mg2+ Stock, #PCR-266"