TopTaq Master Mix Kit

Read phase-out notice below
For highly reliable end-point PCR with unrivaled ease of use

Features

  • Ready-to-use master mix with minimal pipetting steps
  • Fridge storage eliminates thawing time
  • Reliable high-yield PCR performance
  • Convenient room-temperature handling
  • Optimized protocol suitable for all PCR assays

Product Details

The TopTaq Master Mix Kit offers maximum convenience by combining all the benefits of specific TopTaq DNA Polymerase with the advantage of a ready-to-use master mix. It contains TopTaq DNA Polymerase, the unique TopTaq PCR Buffer that minimizes the requirement for optimization, and dNTPs. The unique TopTaq Stabilizer included in the enzyme storage buffer allows storage of TopTaq Master Mix at 4°C and reaction setup at room temperature, without time-consuming thawing of reagents. Providing all components in a ready-to-use master mix reduces pipetting steps, which lowers the risk of contamination. High yields of PCR product are achieved, even after storing the TopTaq Master Mix for extended periods of time at 25°C or 4°C.

 

IMPORTANT NOTE: This product is phasing-out until October 15, 2020, and afterward, available only until stocks last. Visit the product page of the successor kit to view improved features or to request a trial kit.

 

For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource.

Performance

TopTaq Master Mix Kit is used for standard and specialized end-point PCR applications without the need for optimization. High yields of PCR product are achieved, even after storing TopTaq Master Mix for 4 months at 25°C or 4°C, demonstrating the fact that TopTaq Master Mix offers a very robust and reliable amplification system (see figure  Reliable high-yield PCR). TopTaq Stabilizer binds to TopTaq DNA Polymerase, preventing polymerase inactivation during long-term storage. With its balanced potassium and ammonium ions, the unique PCR buffer included in the master mix promotes specific primer–template annealing and simultaneously reduces nonspecific annealing. TopTaq DNA Polymerase outperformed DNA polymerases tested from other suppliers and ensures high yields of specific products, even when using low template amounts, and independent of fragment size (see figure  High PCR product yields without optimization).
See figures

Principle

TopTaq Master Mix Kit has been developed by QIAGEN to provide highly reliable end-point PCR with unrivaled ease of use. Until the development of the TopTaq PCR system, all PCR enzymes required storage at –20°C. Due to the unique proprietary TopTaq Stabilizer contained in the enzyme storage buffer, TopTaq Master Mix Kit is the first PCR kit that can be stored at 4°C. This ready-to-use master mix format ensures greater convenience. High yields of specific products are achieved due to TopTaq DNA Polymerase, even when amplifying fragments of different sizes (see figure " High PCR product yields without optimization").

TopTaq PCR Buffer

The innovative TopTaq PCR Buffer facilitates the amplification of specific PCR products. During the annealing step of every PCR cycle, the buffer greatly increases the ratio of specific primer binding over nonspecific primer binding. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer-annealing conditions over a wider range of temperatures and Mg2+ concentrations than conventional PCR buffers (see figure " Increased specificity of primer annealing"). The need to optimize PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced, or often not required.

TopTaq Stabilizer

TopTaq Stabilizer binds to TopTaq DNA Polymerase, preventing polymerase inactivation during long-term storage (see figure " Stabilization at 4°C"). Template DNA and primers are also prevented from binding to the polymerase at low temperatures. During the initial denaturation step, TopTaq Stabilizer freely dissociates from the polymerase without compromising polymerase activity.  

CoralLoad Concentrate

For enhanced convenience, TopTaq Master Mix Kit includes CoralLoad Concentrate. CoralLoad Concentrate contains a gel-loading reagent and two gel-tracking dyes. This unique reagent improves pipetting visibility and facilitates estimation of DNA migration distance and optimization of agarose gel run time. When using CoralLoad Concentrate, PCR products can be directly loaded onto an agarose gel without prior addition of loading buffer (see figure " Convenient CoralLoad Concentrate").

See figures

Procedure

The TopTaq Master Mix Kit offers all of the benefits of specific TopTaq DNA Polymerase combined with the advantage of a ready-to-use master mix. With the TopTaq Master Mix Kit, separate pipetting of individual components is not required and the risk of contamination and pipetting variability is minimized, making PCR setup quick and easy. During the initial denaturation step, the TopTaq Stabilizer freely dissociates from the polymerase without compromising polymerase activity. This added convenience allows immediate reaction setup without the use of ice — saving the time usually required for reagent thawing (see flowchart " TopTaq procedure"). 
Reliable high-yield PCR without the need for optimization

The optimized protocol suitable for all PCR assays eliminates the need for lengthy optimization of experimental parameters for individual primer–template systems, resulting in substantial time and cost savings. High yields of specific PCR product are achieved, even after storing the TopTaq Master Mix for 4 months at 25°C or 4°C (see figure " Reliable high-yield PCR").

See figures

Applications

The TopTaq Master Mix Kit is highly suited to all end-point PCR applications.

Supporting data and figures

Specifications

FeaturesSpecifications
applicationsPCR, RT-PCR, DNA Fingerprinting
realtimeorendpointEndpoint
dntpsincludedYes (in Master Mix)
mastermixYes
enzymeactivity5' -> 3' exonuclease activity
reactiontypePCR amplification
sampletargettypeGenomic DNA and cDNA
singleormultiplexSingle
storageconditionsFridge storage and room temperature setup
withwithouthotstartWithout Hotstart

Resources

Kit Handbooks (1)
For standard and specialized end-point PCR applications without the need for optimization
Brochures & Guides (3)
Second edition — innovative tools
Addressing critical factors and new solutions

FAQ

Which nucleotide analogs can be used with TopTaq DNA Polymerase?

Nucleotide analogs that can be used with TopTaq DNA Polymerase are:

  • dNTP
  • ddNTP
  • biotin-dATP
  • dITP
FAQ ID -1741
Is TopTaq DNA Polymerase a hot-start polymerase?

No. TopTaq DNA Polymerase is not a hot-start polymerase. Although TopTaq Stabilizer reduces enzyme accessibility at lower temperatures, it does not allow stringent hot-start procedures.

 

FAQ ID -1748
Does TopTaq Stabilizer of the TopTaq DNA Polymerase and Master Mix Kits interfere with downstream applications?

No. TopTaq Stabilizer contained in the enzyme storage buffer of the TopTaq DNA Polymerase and TopTaq Master Mix Kits does not interfere with downstream applications such as cloning, sequencing, or restriction digestion.

 

 

FAQ ID -1744
Do CoralLoad dyes supplied in various QIAGEN PCR Kits interfere with downstream applications?

CoralLoad dyes supplied in PCR Kits such as, e.g., Taq, HotStarTaq, and TopTaq DNA Polymerase and TopTaq Master Mix do not interfere with most downstream enzymatic applications.

However, for reproducible results, purification of PCR products using the QIAquick or MinElute PCR Purification Kits prior to enzymatic manipulation is recommended.

 

 

FAQ ID -1745
Can TopTaq Master Mix simply be stored at room temperature on the lab bench for up to 4 months?

Our data show stability of the TopTaq Master Mix Kit at a constant temperature of 25°C for up to 4 months. However, we do not recommend storage of TopTaq Master Mix on the bench due to temperature fluctuations typical in standard laboratories; nor can we guarantee the performance of TopTaq Master Mix under this storage condition.

FAQ ID -2052
How comparable is CoralLoad gel loading dye contained in various QIAGEN PCR Kits to Sigma Red?

CoralLoad gel tracking dye contained in Taq, HotStarTaq, TopTaq DNA Polymerase and TopTaq Master Mix Kits separates into 2 fragment-size dependent colors (orange and red) when loaded onto an agarose gel. Sigma Red buffer only has one color which is harder to visualize.

 

 

FAQ ID -1644
What is the shelf-life of TopTaq DNA Polymerase and Master Mix?

TopTaq DNA Polymerase and the TopTaq Master Mix Kit are shipped on dry ice but retain full activity at room temperature (15-25°C) for at least 2 weeks. Both kits can be stored at 2-8°C for up to 12 months.

 

 

 

FAQ ID -1743
What is the upper length of PCR products that can be amplified with TopTaq DNA Polymerase and Master Mix Kits?

The maximum amplicon length we have tested using TopTaq DNA Polymerase and the TopTaq Master Mix Kit was 5 kb, amplified from genomic DNA template. Larger PCR amplicon sizes are possible when using less complex template DNA.

 

 

 

 

FAQ ID -1738
What is the fidelity of TopTaq DNA Polymerase?

The error rate of TopTaq DNA Polymerase is very similar to that of standard Taq DNA Polymerase: approximately 2-3x 10e-5 (per base, per cycle).

 

 

 

 

FAQ ID -1739
Can TA/UA cloning be done with PCR products amplified with TopTaq DNA Polmyerase and TopTaq Master Mix Kits?

Yes. The terminal transferase activity of TopTaq DNA Polymerase adds a 3`A-overhang to the PCR product and therefore enables TA- and UA- cloning.

 

 

 

FAQ ID -1740