QIAGEN HotStarTaq Plus DNA Polymerase and Master Mix, and HotStar HiFidelity Polymerase Kit

For highly specific hot-start PCR with minimal optimization.

QIAGEN offers an unrivalled portfolio of highly specific and sensitive hot-start PCR products for a wide range of applications with minimal optimization required.

HotStarTaq Plus DNA Polymerase

• Highly specific amplification with minimal optimization required.
• Fast 5-minute enzyme activation time for increased speed.
• Convenient procedure with ready-to-load PCR buffer and room-temperature reaction setup.

HotStarTaq Plus DNA Polymerase, together with unique CoralLoad or original PCR Buffer (provided), offers the same high specificity and unrivalled sensitivity as HotStarTaq DNA Polymerase, combined with an even faster 5-minute enzyme activation step. Amplification of non-specific products, primer-dimer formation, and background smear are minimized in every PCR cycle. Highly specific results are obtained even when amplifying DNA from single cells.

Figure 1 Description: Gel electrophoresis image comparing the specificity of HotStarTaq Plus DNA Polymerase against other hot-start PCR enzymes and standard Taq DNA Polymerase. Lanes labeled 'M' show molecular weight markers. The results indicate that QIAGEN's HotStarTaq Plus and HotStarTaq enzymes produce a single, sharp band of the expected size (1.5 kb fragment of the human CFTR gene), while other suppliers' enzymes show non-specific products, primer-dimers, or background smears.

Figure 2 Description: Gel electrophoresis illustrating the high sensitivity and specificity of HotStarTaq and HotStarTaq Plus DNA Polymerases in single-cell PCR. The gel image shows amplified DNA fragments from a 500 bp fragment of the murine p53 gene. Both QIAGEN enzymes produced clear, specific bands, demonstrating their effectiveness even with minimal template DNA.

Convenient kit format

The kit comes complete with enzyme, original PCR buffer, and ready-to-load CoralLoad PCR Buffer, providing specific amplification with minimal optimization required. Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates, is also provided.

HotStarTaq Plus Master Mix Kit

The HotStarTaq Plus Master Mix Kit provides all of the benefits of HotStarTaq Plus DNA Polymerase and more, including:

• Fewer pipetting steps, reducing the risk of contamination and pipetting error.
• All components in one kit, including dNTPs.

Fast, easy, and convenient PCR setup

The ready-to-use master mix contains HotStarTaq Plus DNA Polymerase, QIAGEN PCR Buffer, dNTPs, and MgCl₂. The kit is also supplied with CoralLoad Concentrate, which can be optionally added to the master mix prior to the start of PCR. With the HotStarTaq Plus Master Mix Kit, there is no need for separate pipetting of individual components, minimizing the risk of contamination. Lengthy calculations are avoided, and setting up amplification reactions is fast and easy. Since errors due to pipetting variability and miscalculations are reduced, PCR results are highly reproducible. The hot start provided by HotStarTaq Plus DNA Polymerase in the Master Mix Kit leads to increased PCR specificity, and amplification reactions can be set up at room temperature.

Minimal PCR optimization with unique PCR Buffer

QIAGEN PCR Buffer has been developed to eliminate the need for optimization of individual primer-template systems, saving time and money. The balanced combination of KCl and (NH₄)₂SO₄ in the buffer promotes specific primer-template annealing. Simultaneously, non-specific annealing is reduced, maximizing yields of specific PCR product. Specific amplification is maintained over a wide range of temperatures and Mg²⁺ concentrations, without the need for time-consuming optimization.

Figure 4 Description: Diagram explaining the mechanism of the unique QIAGEN PCR Buffer. It depicts how K⁺ ions bind to the phosphate groups on the DNA backbone, stabilizing primer annealing to the template. Simultaneously, NH₄⁺ ions interact with base pairing, destabilizing weak hydrogen bonds at mismatched bases. This dual action is shown to maintain a high ratio of specific to non-specific primer-template binding across a range of temperatures.

Convenient ready-to-load PCR buffer

The novel CoralLoad PCR Buffer provided with HotStarTaq Plus DNA Polymerase and CoralLoad Concentrate provided with HotStarTaq Plus Master Mix contains two gel-tracking dyes, enabling immediate loading of your PCR product onto an agarose gel after amplification. Movement of the DNA on the gel can be easily monitored using the gel-tracking dyes.

Figure 3 Description: Images of a PCR tube containing CoralLoad PCR Buffer and wells on an agarose gel. CoralLoad PCR Buffer is described as containing two gel-tracking dyes that allow for immediate loading of PCR products onto an agarose gel and easy visualization of DNA migration.

HotStar HiFidelity Polymerase Kit

The HotStar HiFidelity Polymerase Kit provides:

• High sensitivity due to novel buffer additive.
• Direct cloning in TA- or UA-cloning vectors.
• High-fidelity amplification, tenfold higher than Taq DNA Polymerase.

High sensitivity and reliability

HotStar HiFidelity DNA Polymerase is a hot-start proofreading enzyme uniquely modified to prevent degradation of primers and template during PCR setup, providing sensitive and specific high-fidelity PCR. The unique buffer system contains a balanced combination of K⁺ and NH₄⁺ ions and includes Factor SB (patent pending), which greatly improves PCR sensitivity. The kit comes complete with enzyme and PCR buffer containing dNTPs and is ready to use with minimal optimization required. Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates, is also provided.

Figure 5 Description: Gel electrophoresis comparison of HotStar HiFidelity DNA Polymerase with four other high-fidelity PCR enzymes. The experiment amplified a 2.3 kb fragment of the human IL9R gene from varying amounts of genomic DNA. The gel shows that QIAGEN's HotStar HiFidelity Polymerase produces a clean, specific band, indicating high sensitivity and reliability, with minimal primer-dimers compared to some other enzymes.

Unique UA/TA cloning facility

In contrast to all other proofreading enzymes, HotStar HiFidelity Polymerase is uniquely modified to produce A overhangs, enabling streamlined and reliable UA/TA cloning. The cloning efficiency is comparable with fragments derived from Taq DNA polymerases, removing the need for less efficient blunt end cloning and time-consuming verification sequencing.

Figure 6 Description: A bar chart comparing cloning efficiency for TA cloning and UA cloning using standard Taq DNA Polymerase versus HotStar HiFidelity Polymerase. The chart shows that HotStar HiFidelity Polymerase achieves significantly higher cloning efficiency, particularly for UA cloning, compared to Taq DNA Polymerase.

Ideal for highly sensitive applications

The HotStar HiFidelity Polymerase Kit is ideal for all hot-start PCR applications that require high sensitivity combined with a low error rate, such as cloning and site-directed mutagenesis. The kit comes complete and ready to use with enzyme, PCR buffer (containing dNTPs), and Q-Solution.

Features of QIAGEN hot-start enzymes

Ordering Information

* Contains 15 mM MgCl₂.

† Larger kit sizes available, please inquire.

‡ Contains 3 mM MgCl₂ and 400 µM each dNTP.

§ Contains Factor SB, dNTPs, and optimized concentration of MgSO₄.

For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at www.qiagen.com or can be requested from QIAGEN Technical Services or your local distributor.

Find out more about QIAGEN's highly-specific hot-start PCR products at www.qiagen.com/goto/PCR.