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 reproducibleÂ 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.
|#2001S||TaqÂ 2x PCR Master Mix||100 reactions Ã¡ 50 Âµl, 2Â x 1.25 ml||Â â‚¬ 45,-|
|#2001M||TaqÂ 2x PCR Master Mix||500 reactions Ã¡ 50 Âµl, 10Â x 1.25 ml||Â â‚¬ 200,-|
Bulk offer: Should you require larger amounts (>1000 reactions), significant discounts are available. Please contact us to receive an individual quotation.
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.
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 2x Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.
|Taq 2x PCR Master Mix||25 Âµl||1x|
|Primer forward (10 ÂµM)*||1 Âµl||0.2 ÂµM (0.05-1 ÂµM)|
|Primer reserve(10 ÂµM)*||1 Âµl||0.2 ÂµM (0.05-1 ÂµM)|
|Template/Sample extract**||x Âµl||Â|
|Nuclease-free water||up to 50Âµl total reaction volume||Â|
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 Âµl (gnomic 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||95Â°C||2 min|
|Annealing*||54-72Â°C||30 secÂ Â Â (25-40 cycles)|
|Extension||72Â Â°C||1 min / 1000 bp|
*Typically, the annealing temperature is about 3-5Â°C below the calculated melting temperature of the primers used.Â
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 andÂ analyzedÂ byÂ 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 -20Â°C.
Minimize the number of freeze-thaw cycles by storing in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4Â°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.
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 TaqÂ 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):6427â€“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): 6008â€“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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