HotStarTaq Plus DNA Polymerase

For fast and highly specific amplification in all applications
  • High PCR specificity with minimal optimization
  • Fast 5-minute enzyme activation time
  • Ready-to-load PCR buffer for faster and easier handling
The polymerase combines the high specificity, sensitivity, and minimal optimization of HotStarTaq DNA Polymerase, with a fast 5-minute activation time. PCR can be set up at room temperature and reactions can be directly loaded onto a gel, due to novel CoralLoad PCR Buffer, which contains two gel-tracking dyes. The standard QIAGEN PCR Buffer is also included for greater convenience. In addition, Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC rich) templates, is also provided. The unique kit components and optimized protocols streamline the PCR procedure.
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HotStarTaq Plus DNA Polymerase (50)
50 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
203601
$37.10
HotStarTaq Plus DNA Polymerase (250)
250 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
203603
$186.00
HotStarTaq Plus DNA Polymerase (1000)
1000 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
203605
$659.00
HotStarTaq Plus DNA Polymerase (5000)
5000 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
203607
$2,987.00
HotStarTaq Plus DNA Polymerase (25000)
25,000 units HotStarTaq Plus DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
203609
$13,399.00
The HotStarTaq Plus DNA Polymerase is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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CoralLoad PCR Buffer.|HotStarTaq Plus procedure.|Highest specificity.|Amplification of difficult templates.|Higher specificity with different primer–template systems.|Effect of hot start on RT-PCR performance.|Highly sensitive single-cell PCR.|Increased specificity of primer annealing.|
[A] CoralLoad PCR Buffer contains 2 gel-tracking dyes for improved pipetting visibility during PCR setup, enabling immediate gel loading of PCR products for [B] easy visualization of DNA migration.|The HotStarTaq Plus procedure is fast and easy for maximum convenience.|PCR was performed with HotStarTaq Plus DNA Polymerase, HotStarTaq DNA Polymerase, and Taq DNA Polymerase from QIAGEN, and three hot-start PCR enzymes from the indicated suppliers. Parallel reactions were performed following the suppliers' recommendations, using 50 ng human genomic DNA. A 1.5 kb fragment of the human CFTR gene was amplified in 35 PCR cycles. M: markers.|Two different primer–template systems were amplified in duplicate using QIAGEN PCR Buffer and Taq DNA Polymerase in the absence () or presence (+) of 1x Q-Solution. Q-Solution enables specific amplification of difficult templates. [A] human angiotensin receptor II gene; [B] mouse protein kinase C gene; M: markers.|Three different primer–template systems were amplified under the same conditions with either Taq DNA polymerase from Supplier R (R) or with HotStarTaq DNA Polymerase (H). System 1: A 1.1 kb fragment of a D-IgI homolog was amplified from human genomic DNA. System 2: A 296 bp fragment from the chromosomal region correlated with X-linked juvenile retinoschisis was amplified from human genomic DNA. System 3: A 214 bp fragment of the β-actin gene was amplified from cDNA synthesized from total RNA. M: markers. Note: Data are shown for HotStarTaq DNA Polymerase; identical sensitivity and specificity were obtained with HotStarTaq Plus DNA Polymerase.|A 1.1 kb fragment of the human interleukin 1 receptor (type II) gene was amplified from cDNA. Amplification reactions were prepared in triplicate using Taq DNA polymerase and buffer from Supplier L (No hot start); antibody-mediated hot start using enzyme and buffer from Supplier L (Antibody-mediated); and HotStarTaq DNA Polymerase and PCR Buffer from QIAGEN (HotStarTaq). M: markers. Note: Data are shown for HotStarTaq DNA Polymerase; identical sensitivity and specificity were obtained with HotStarTaq Plus DNA Polymerase.|A 500 bp fragment of the murine p53 gene was amplified from single cells isolated by flow cytometry, and directly sorted into individual PCR tubes. Reactions were prepared in parallel using HotStarTaq DNA Polymerase and PCR Buffer from QIAGEN (HotStarTaq), a hot-start enzyme and buffer from Supplier AII (Hot-start enzyme), or antibody-mediated hot start and buffer from Supplier L (Antibody-mediated). M: markers. Note: Data are shown for HotStarTaq DNA Polymerase; identical sensitivity and specificity were obtained with HotStarTaq Plus DNA Polymerase.|Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K+ binds to the phosphate groups (P) on the DNA backbone, stabilizing the annealing of the primers to the template. NH4+, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing the weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains a high ratio of specific-to-nonspecific primer-template binding over a wide temperature range.|
Performance
Each lot of HotStarTaq Plus DNA Polymerase is subjected to a comprehensive range of quality control tests, including a stringent PCR specificity and reproducibility assay in which low-copy targets are amplified. HotStarTaq Plus DNA Polymerase outperformed kits tested from other suppliers and ensures high specificity and superior performance in hot-start PCR (see figures "Highest specificity" and "Higher specificity with different primer–template systems", and table). The innovative PCR buffer provided with the kit ensures specificity over a wide range of PCR conditions, minimizing the need for optimization. Suboptimal PCR can be improved with Q-Solution, also provided with the kit (see figure "Amplification of difficult templates"). Together, these components ensure specific amplification in a range of applications (see figure "Effect of hot start on RT-PCR performance" and "Highly sensitive single-cell PCR").

Comparison of hot-start methods 
HotStarTaq Plus DNA Polymerase HotStarTaq DNA Polymerase Hot-start enzyme from Supplier AII Supplier R Supplier I (antibody-mediated) Manual Wax  barrier
Specific amplification ++ ++ + ++ + +/– +/–
Minimal PCR optimization ++ ++ +/– +/– +/–
Easy to use +++ ++ ++ + +
Speed of activation ++ + ++ ++ ++


HotStarTaq Plus DNA Polymerase specifications

Concentration: 5 units/µl
Recombinant enzyme: Yes
Substrate analogs: dNTP, ddNTP, dUTP, biotin-11-dUTP, DIG-11-dUTP, fluorescent-dNTP/ddNTP
Extension rate: 2–4 kb/min at 72°C
Half-life: 10 min at 97°C ; 60 min at 94°C
Amplification efficiency: ≥105 fold
5'–>3' exonuclease activity: Yes
Extra A addition: Yes
3'–>5' exonuclease activity: No
Contaminating nucleases: No
Contaminating RNases: No
Contaminating proteases: No
Self-priming activity: No

Principle

HotStarTaq Plus DNA Polymerase provides the unrivaled performance of HotStarTaq DNA Polymerase with a shortened activation time of just 5 minutes.

HotStarTaq Plus DNA Polymerase, a modified form of QIAGEN Taq DNA Polymerase, is supplied in an inactive state that has no polymerase activity at ambient temperatures. This prevents extension of nonspecifically annealed primers and primer dimers formed at low temperatures during PCR setup and the initial PCR cycle (see figures "Highest specificity" and "Higher specificity with different primer-template systems"). HotStarTaq Plus DNA Polymerase is activated by a short 5-minute incubation at 95°C which can be easily incorporated into any existing thermal-cycler program.

QIAGEN PCR Buffer

QIAGEN PCR Buffer maintains specific amplification in every cycle of PCR by promoting a high ratio of specific-to-nonspecific primer binding during the annealing step in each PCR cycle (see figure "Increased specificity of primer annealing"). Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the buffer provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is therefore often minimal or not required. 

CoralLoad PCR Buffer

HotStarTaq Plus DNA Polymerase is supplied with CoralLoad PCR Buffer, which has all of the advantages of QIAGEN PCR Buffer but can also be used to directly load the PCR reaction onto an agarose gel without the need to add a gel loading buffer. CoralLoad PCR Buffer provides the same high PCR specificity and minimal reaction optimization as the conventional QIAGEN PCR Buffer. Additionally, it contains two marker dyes — an orange dye and a red dye — that facilitate estimation of DNA migration distance and optimization of agarose gel run time (see figure "CoralLoad PCR Buffer"). The buffer ensures improved pipetting visibility and enables direct loading of PCR products onto a gel, for enhanced convenience.  

Q-Solution

Q-Solution, an innovative PCR additive that facilitates amplification of difficult templates by modifying the melting behavior of DNA, is also provided with HotStarTaq DNA Polymerase. This unique reagent improves suboptimal PCR caused by templates that have a high degree of secondary structure or GC-rich templates (see figure "Amplification of difficult templates"). Unlike other commonly used PCR additives such as DMSO, Q-Solution is used at just one working concentration, is nontoxic, and PCR purity is guaranteed. Adding Q-Solution to the PCR does not compromise PCR fidelity.

Procedure
HotStarTaq Plus DNA Polymerase is supplied with a streamlined, optimized protocol for fast and easy PCR setup. HotStarTaq Plus DNA Polymerase is activated by a 5-minute, 95°C incubation step, which can easily be incorporated into existing thermal cycling programs. Reactions can be set up at room temperature, ensuring greater convenience and ease of use (see figure "HotStarTaq Plus procedure"). CoralLoad PCR Buffer, also provided with the kit, ensures improved pipetting visibility and enables direct loading of PCR products onto a gel for enhanced convenience.
Applications

HotStarTaq Plus DNA Polymerase is highly suitable for a wide variety of applications, including challenging applications such as amplification of:

  • Complex genomic templates
  • Complex cDNA templates (e.g., RT-PCR)
  • Very low-copy targets (e.g., single-cell PCR)
  • Reactions with multiple primer pairs
Feature
Specifications
Applications PCR, RT-PCR, Complex genomic templates, very low-copy targets
Enzyme activity 5' -> 3' exonuclease activity
Mastermix No
Reaction type PCR amplification
Real-time or endpoint Endpoint
Sample/target type Genomic DNA and cDNA
Single or multiplex Single
With/without hotstart With hotstart

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Kit Handbooks
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For highly specific hot-start PCR without optimization  
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至適化不要で特異性の高いホットスタートPCR
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