Taq Polymerase

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:
The polymerase is recommended for routine PCR applications (up to 4 kb fragment length) or high throughput PCR.
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:
Reaction Buffer and Nucleotides are not included. Please refer to our sections Buffers & Components and Nucleotides.

Taq Polymerase
5 units/μl Taq DNA Polymerase in 20 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizers, 50 % (v/v) Glycerol, pH 8.0 (25°C)

Assay Set-Up:
Before starting, thaw up all components and vortex thoroughly to ensure homogeneity.

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

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.

Recommended Buffer Systems:
Ruby Buffer (#PCR-272) includes a density reagent + tracking dye and allows the direct loading of the PCR products into an electrophoresis gel. For DNA detection / fluorescent DNA staining, we recommend to use new generations dyes (e.g. SYBR 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 (e.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.,

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.,

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 inaccuracy.

Optimization of MgCl₂ concentration:
MgCl₂ Solution - 25 mM (#PCR-266) is recommended for optimization of the final Mg²⁺ concentration. A concentration of 1.5-2.0 mM is required for optimal functionality. Lower concentrations give higher specificity, whereas higher concentrations give higher yield.

Related products:

• dNTP Mix - 10 mM, #NU-1023
• Ruby Buffer, #PCR-272
• Crystal Buffer, #PCR-271
• KCl Buffer, #PCR-262
• MgCl₂ Solution - 25 mM, #PCR-266"