DNA / PCR

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.

 

ArticleProductSizePrice
#1001 STaq DNA polymerase400 U, 5U/µl, 80 µl € 45,-
#1001 MTaq DNA polymerase1000 U, 5U/µl, 200 µl € 105,- 
#1001 LTaq DNA polymerase4000 U, 5U/µl, 2x 400 µl € 400,-
#1101 STaq DNA polymerase hot-start400 U, 5U/µl, 80 µl € 65,-
#1101 MTaq DNA polymerase hot-start1000 U, 5U/µl, 200 µl € 145,-
#1101 LTaq DNA polymerase hot-start4000 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.

 

download-pdf Product manual (PDF) Order

 

 

Example Results

Example Results

Faster Detection and Higher Sensitivity

 

high_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

easy-taq-2x-mastermix-1

 

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.

 

Reaction Setup

Reaction Setup

ComponentVolumeFinal concentration
Primer forward (10 µM)*1 µl0.2 µM (0.05-1 µM)
Primer reverse(10 µM)*1 µl0.2 µM (0.05-1 µM)
dNTPs (2 mM)5 µl200 µM
10 x reaction buffer5 µl1x
Taq DNA polymerase 5 U/µl0.25 µl1.25 U / reaction
Template/Sample extract**x µl 
Nuclease-free waterup 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).

Applications

Applications

  • 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

StepTemperatureTime
Initial denaturation95°C2 min
Denaturation95°C15 sec
Annealing*54-72°C30 sec     (25-40 cycles)
Extension72 °C1 min / 1000 bp
Hold<10°Chold

*Typically, the annealing temperature is about 3-5°C below the calculated melting temperature of the primers used. 

 

 

Quality Control

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 info@mypols.de for the lot-specific concentration.
No contamination has been detected in standard test reactions.

 

Storage

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.

 

easy-taq-2x-mastermix-textimg

 

img-faqsFAQ`s

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.


img-contactContact & Support

img-accordionContact for support

img-accordionNeed project consulting?

img-accordionPublish and Get A Free Kit

 

img-downloadDownload

img-accordionMSDS (Material Safety Data Sheet)

img-accordionProduct manual

img-accordionOrder form

 

 

Comments are closed.