DNA / PCR

Taq 2x PCR Master Mix

 

is a ready to use reaction mix. It contains all components necessary for a successful and reliable PCR or primer extension reaction in a standard PCR cycler including Taq wild-type DNA polymerase and ultrapure dNTPs. Only primers and template need to be added. This mix provides robust PCR performance for a wide range of PCR applications. The pre-ready 2x mix ensures reproducible results, significantly reduces set-up times and the risk of pipetting mistakes. It can also be used for realtime cycling, when adding a suitable realtime dye or a fluorescent probe.

 

Article Product Size Price
#2001S Taq 2x PCR Master Mix 100 reactions á 50 µl, 2 x 1.25 ml  € 45,-
#2001M Taq 2x PCR Master Mix 500 reactions á 50 µl, 10 x 1.25 ml  € 200,-

Bulk offer: Should you require larger amounts (>1000 reactions), significant discounts are available. Please contact us to receive an individual quotation.

 

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Example Results

Example Results

Broad Amplification Range

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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.

 

 

 

 

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 2x Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.

 

Reaction Setup

Reaction Setup

Component Volume Final concentration
Taq 2x PCR Master Mix 25 µl 1x
Primer forward (10 µM)* 1 µl 0.2 µM (0.05-1 µM)
Primer reserve(10 µM)* 1 µl 0.2 µM (0.05-1 µM)
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 (gnomic 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

Step Temperature Time
Initial denaturation 95°C 2 min
Denaturation 95°C 15 sec
Annealing* 54-72°C 30 sec     (25-40 cycles)
Extension 72 °C 1 min / 1000 bp
Hold <10°C hold

*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 Mastermix 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.
Minimize the number of freeze-thaw cycles by storing in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C.

 

Composition / Content of the Product

Taq 2x PCR Master Mix is containing all the components necessary for PCR, including wild-type Taq DNA polymerase and an optimized buffer including ultrapure dNTPs.

 

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.


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