Taq 2x PCR Master Mix

Product#: 2001M
$320.00
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Taq 2x PCR Master Mix

Cat. No. 2001M
Size 500 tests
Store  -20°C, This product is shipped on cool packs. It is recommended to store the product upon arrival at -20°C.

Description

Taq 2x PCR Master Mix - ready to use mix simplifies your PCR setup. Only primers and template need to be added as the mix contains all copmponents for a successful and reliable PCR. This ensures reproducible results, significantly reduces set-up times and the risk of pipetting errors. The robustness of PCR performance allows the application of this mix in a wide range of PCR amplifications.

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 all standard PCR cyclers. 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.
Contents
This mix contains a Taq DNA polymerase variant in reaction buffer and ultrapure dNTPs. The DNA polymerase is hotstart-formulated with an aptamer. Temperatures above 50-55°C cause the aptamer’s secondary structure to melt and will set-free the DNA polymerase. It can also be used for real-time cycling, when adding a suitable realtime dye, for example GreenDye (#2000), or a fluorescent probe.

Taq 2x PCR Master Mix contains all the components necessary for PCR, including Taq DNA polymerase variant and an optimized buffer including ultrapure dNTPs
Applications
  • Standard PCR
  • Realtime-PCR (addition of suitable dye required)
  • Primer extension reactions
  • TA cloning
  • 3’A-tailing of blunt ends
  • Screening / High-throughput PCRs
Recommendations for PCR/ Reaction Setup
PCR Mix

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 reverse (10 µM)*

1  µl

0.2 µM (0.05-1 µM)

Template/Sample extract

x  µl

<1000 ng** DNA

Nuclease-free  water

up to 50µl total volume

* Primers should ideally have a GC content of 40-60% typically
** Suggested template concentration should be about 1 ng - 1 µg (genomic DNA) or 1 ng – 1 pg (plasmid/viral DNA).

Typical 3-step PCR protocol

Initial denaturation

95°C

2 min

Denaturation

95°C

15 sec

Annealing

54–72°C

30 sec

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Extension

72°C

1 min/1000 bp

25-40 cycles

Hold

<10°C

 

 

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

Recommendations for sample handling
  • Keep all components on ice.
  • Spin down and mix all solutions carefully before use.
  • Primers should ideally have a GC content of 40-60%.
  • Suggested template concentration should be about 1 ng – 1 pg (plasmid/viral DNA) or 1 ng - 1 µg (genomic DNA).
  • 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.
Quality Control Assays
Taq 2x PCR Master Mix is tested for successful PCR performance. A 92 bp fragment (beta-actin gene) was amplified from human genomic DNA and analysed by agarose gel electrophoresis. The activity of Taq DNA polymerase has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and DNA primer. Enzyme-concentration has been determined by proteinspecific staining. Please inquire more information at info@mypols.de for the lot-specific concentration. No contamination has been detected in standard test reactions.
Material Safety Data (MSDS)
According to OSHA 29CFR1910.1200, Australia [NOHSC:1005, 1008 (1999)] and the EU Directives 67/548/EC, 1999/45/EC and 1272/2008 (CLP Regulation) any products which do not contain more than 1% of a component classified as dangerous or hazardous nor more than 0.1% of a component classified as carcinogenic, do not require a MSDS. However, we recommend the use of gloves, lab coats and eye protection when working with these or any other chemical reagents. myPOLS Biotec takes no liability for damage resulting from handling or contact with this product. This product is not hazardous, not toxic, not IATA-restricted. Product is not from human, animal or plant origin. The source of the product is recombinant protein expression in E. coli. The product is for research use only and may be used for in-vitro experiments only.
References
  1. 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.
  2. Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry 1988; 27(16): 6008–6013. K. R. Tindall, T. A. Kunkel.
  3. 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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