Taq DNA polymerase (hotstart)
Taq DNA polymerase
is supplied together with the 10x Taq reaction buffer. The reaction buffer has been specifically designed for optimal PCR performance and polymerase activity. Taq DNA polymerase can also be used for realtime cycling, when adding a suitable realtime dye or a fluorescent probe. Optionally Taq DNA polymerase is also available as hot-start formulation that prevents non-specific amplification during the reaction set-up.
|#1101 L||Taq DNA polymerase hot-start||4000 U, 5U/µl, 2x 400 µl||€ 520,-|
Bulk offer: Should you require larger amounts (> 10.000 U), significant discounts are available. Please contact us to receive an individual quotation.
Faster Detection and Higher Sensitivity
A fragment (64 bp) of the human blood-coagulation factor IIa (F2) was amplified from 20 ng, 2 ng, 200 pg and 20 pg of a human genomic DNA extract. The same PCRs were performed in parallel using the Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.
Broad Amplification Range
Different-sized amplicons from 3 ng of a DNA plasmid were amplified. The use of Taq 2x PCR Master Mix resulted in clean and high yield of products, as analysed after PCR on a 0.8% agarose gel.
|Primer forward (10 µM)*||1 µl||0.2 µM (0.05-1 µM)|
|Primer reverse(10 µM)*||1 µl||0.2 µM (0.05-1 µM)|
|dNTPs (2 mM)||5 µl||200 µM|
|10 x reaction buffer||5 µl||1x|
|Taq DNA polymerase 5 U/µl||0.25 µl||1.25 U / reaction|
|Template/Sample extract**||x µl|
|Nuclease-free water||up to 50µl total reaction volume|
Keep all components on ice.
Spin down and mix all solutions carefully before use.
* Primers should ideally habe a GC-content of 40-60%
**Suggested template concentration should be about 1 ng – 1 µl (genomic DNA) or 1 ng – 1 pg (plasmid/viral DNA).
- Standard PCR
- Realtime PCR
- Primer extension reaction
- TA cloning
- 3`A-tailing of blunt ends
- Screening / Hight-throughput PCRs
Typical PCR protocol
Typical PCR protocol
|Initial denaturation||95°C||2 min|
|Annealing*||54-72°C||30 sec (25-40 cycles)|
|Extension||72 °C||1 min / 1000 bp|
*Typically, the annealing temperature is about 3-5°C below the calculated melting temperature of the primers used.
PCR activity: Taq DNA polymerase was tested for successful PCR performance. A 92 bp fragment (beta-actin gene) was amplified from human genomic DNA and analyzed by agarose gel electrophoresis.
DNA polymerase activity: Taq DNA polymerase activity has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer.
Enzyme-concentration has been determined by protein-specific staining. Please inquire more information at firstname.lastname@example.org for the lot-specific concentration.
No contamination has been detected in standard test reactions.
This product is shipped on cool packs. Please store the product upon arrival at -20°C.
Composition / Content of the Product
Taq DNA polymerase is supplied as a 5 U/µl solution containing glycerol. It comes together with a 10x optimized reaction buffer.
How can I optimize the PCR reaction conditions?
1. The annealing temperature can usually be optimized. Try a temperature gradient and determine the best annealing temperature, which yields in the cleanest product.
2. Add a gradual amount of betaine 0-1 M or DMSO 0-7.5% to the reaction mix and select for the cleanest product and the highest yield.
3. Try to shorten the extension and annealing time. Too long and too many cycles may lead to over-amplification and side-products.
Where can I read more about the Taq DNA polymerase?
Have a look under following references for example:
Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus. J Biol Chem. 1989; 264(11):6427–6437. F. C. Lawyer, S. Stoffel, R. K. Saiki, K. Myambo, R. Drummond, and D. H. Gelfand.
Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase.
Biochemistry 1988; 27(16): 6008–6013. K. R. Tindall, T. A. Kunkel.
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
Science 1988; 239(4839): 487-491. R. K. Saiki, D. H. Gelfand, S. Stoffel, S. J. Scharf, R. Higuchi, G. T. Horn, K. B. Mullis and H. A. Erlich.
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