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  • Rapid, high-performance multiplex PCR
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standard pcr Hot-start Takara Taq for research use
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standard pcr Takara Taq
standard pcr Hot-start Takara Taq for research use
Tech Note

Highly efficient and specific multiplex PCR using TaKaRa Taq DNA Polymerase Hot Start Version

The hot-start (HS) version of TaKaRa Taq DNA Polymerase provides higher efficiency and specificity than the standard TaKaRa Taq DNA Polymerase when used in multiplex PCR, with an efficiency comparable to individual amplification reactions.

Introduction Results Conclusions Methods

Introduction  

Multiplex PCR is a variant of traditional PCR which allows simultaneous amplification of numerous targets using multiple primer pairs in a single amplification reaction. This approach is used in many areas of scientific research, including genotyping applications that require analysis of multiple markers. It is also used in pathogen detection and in analysis of polymorphisms. The technique is demanding, and successful multiplex amplifications require careful primer design; reaction optimization; and a specific, sensitive, and reliable DNA polymerase.

TaKaRa Taq DNA Polymerase Hot Start Version contains a mixture of high-purity TaKaRa Taq DNA Polymerase and a monoclonal antibody to Taq DNA polymerase, which binds to the polymerase until the reaction temperature is elevated. The binding of this antibody prevents nonspecific amplification due to mispriming and/or formation of primer dimers during reaction assembly. The antibody is then denatured in the initial PCR DNA-denaturation step, releasing the polymerase and allowing DNA synthesis to proceed. Takara HS Taq provides sensitive, reliable amplification, along with reduced background and increased reaction specificity and efficiency.

Results  

Comparison with a standard Taq DNA polymerase

The performance of Takara HS Taq in a multiplex reaction was compared to its performance in single-target reactions, and to that of the standard TaKaRa Taq DNA Polymerase in a multiplex reaction (Figure 1). Takara Taq and Takara HS Taq were each used to amplify a human genomic DNA template with eight different primer pairs, each specific for a target ranging from 84 to 432 bp in size. In addition, Takara HS Taq was used to perform an individual amplification reaction with each of the eight primer pairs.

The sample amplified using Takara HS Taq (Lane 10) shows more efficient multiplex amplification of all the bands individually amplified in Lanes 1–8 than the sample amplified using Takara Taq (Lane 9), and it lacks the low molecular weight non-specific amplification band seen with Takara Taq (Lane 9). The intensities of the bands in Lane 10 are comparable to the intensities of the corresponding individual bands in Lanes 1–8, indicating that Takara HS Taq provides multiplex efficiencies comparable to those observed in individual amplification reactions.

Comparison of TaKaRa Taq Hot Start to standard TaKaRa Taq

Comparing the ability of TaKaRa Taq DNA Polymerase Hot Start Version to amplify human genomic DNA fragments to that of the standard TaKaRa Taq DNA Polymerase. PCR reactions were performed using human genomic DNA as a template and primer pairs for eight different targets. Lanes 1–8 contain individual reactions for each primer pair amplified using Takara HS Taq. Lanes 9 and 10 contain multiplex PCR reactions performed with all eight primer pairs in a single tube, amplified with either the standard Taq DNA polymerase (Takara Taq, Lane 9) or Takara HS Taq (Lane 10).

Conclusions  

This experiment demonstrates that target amplification efficiencies for multiplex PCR using Takara HS Taq are comparable to efficiencies observed for separate (single target) amplification reactions. In addition, Takara HS Taq demonstrates superior efficiency and specificity over standard Taq polymerase (Takara Taq) in this multiplex-PCR application.

Methods  

PCR cycling conditions for individual reactions

1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72°C for 60 sec.

PCR cycling conditions for multiplex reactions

1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 57°C for 30 sec, and 72°C for 60 sec, followed by a final step at 72°C for 90 sec.

Learn more about TaKaRa Taq DNA Polymerase Hot Start Version »

Related products

Cat. # Product Size Price License Quantity Details
R007A TaKaRa Taq™ DNA Polymerase Hot Start Version 250 Units USD $195.00

An antibody-mediated hot-start version of TaKaRa Taq DNA Polymerase, which is a recombinant version of full-length Taq polymerase. It has the same characteristics and capabilities as the native Taq polymerase, and is suitable for a variety of standard PCR applications. The polymerase is supplied with separate tubes of buffer (Mg2+ plus) and dNTPs.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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R007A: TaKaRa Taq DNA Polymerase Hot Start Version

R007A: TaKaRa Taq DNA Polymerase Hot Start Version
R007B TaKaRa Taq™ DNA Polymerase Hot Start Version 1,000 Units USD $702.00

An antibody-mediated hot-start version of TaKaRa Taq DNA Polymerase, which is a recombinant version of full-length Taq polymerase. It has the same characteristics and capabilities as the native Taq polymerase, and is suitable for a variety of standard PCR applications. The polymerase is supplied with separate tubes of buffer (Mg2+ plus) and dNTPs. Cat. # R007B contains 4 of Cat. # R007A. Please refer to Cat. # R007A for complete product documentation and resources.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Image Data

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R007B: TaKaRa Taq DNA Polymerase Hot Start Version

R007B: TaKaRa Taq DNA Polymerase Hot Start Version
R028A Premix Taq™ DNA Polymerase Hot-Start Version 100 Rxns USD $185.00

A convenient 2X PCR master mix which consist of Takara Taq HS polymerase, optimized reaction buffer, and dNTPs. Takara Taq HS polymerase is an antibody-mediated hot-start version of TaKaRa Taq DNA Polymerase, which is a recombinant version of full-length Taq polymerase. Takara Taq has the same characteristics and capabilities as the native Taq polymerase, and is suitable for a variety of standard PCR applications.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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R028A: Premix Taq DNA Polymerase Hot-Start Version

R028A: Premix Taq DNA Polymerase Hot-Start Version
R300A TaKaRa Taq™ HS Perfect Mix 100 Rxns USD $147.00

TaKaRa Taq HS Perfect Mix is a convenient, hot-start 2X PCR master mix that includes DNA polymerase, optimized reaction buffer, and dNTPs. Perfect Mix contains a modified Taq DNA polymerase, which lacks exonuclease activities. It is designed for rapid extension and good PCR specificity.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

R300A: TaKaRa Taq HS Perfect Mix

R300A: TaKaRa Taq HS Perfect Mix
R300B TaKaRa Taq™ HS Perfect Mix 400 Rxns USD $585.00

TaKaRa Taq HS Perfect Mix is a convenient, hot-start 2X PCR master mix that includes DNA polymerase, optimized reaction buffer, and dNTPs. Perfect Mix contains a modified Taq DNA polymerase, which lacks exonuclease activities. It is designed for rapid extension and good PCR specificity. Cat. # R300B contains 4 of Cat. # R003A. Please refer to Cat. # R300A for complete product documentation and resources.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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