Reagents for PCR - Selection Guide

> Enzymes for Routine and Hot Start
> Enzymes for Specific Requirements

Enzymes for Routine and Hot Start

Enzymes for Specific Requirements

* Requiring carryover prevention and 6x higher fidelity? Choose Expand High Fidelity^PLUS PCR System.
** For amplicon sizes of 20-35 kb choose Expand 20 kb^PLUS PCR System.

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Products for Conventional PCR

In this section of the website we present general information on Roche Applied Science amplification products. To explore this information, use the links below.
 

> Enzymes for PCR
> PCR Enzyme dNTPacks
> Mastermixes and kits for PCR

Related Products for Conventional PCR
 

PCR is only one part of a sequence of working steps that usually begins with the preparation of a nucleic acid (DNA, RNA), sometimes followed by reverse transcription prior to the PCR. Depending on your application, there are many downstream steps after PCR including, post-PCR purification, cloning, labeling or analyzing DNA.  Roche Applied Science not only provides outstanding products for the PCR step, but also offers many reagents, kits, and other tools that can greatly facilitate your work.
 

• PCR-grade nucleotides
• Mastermixes for Real-Time PCR
• Products for one step RT-PCR
• Products for two step RT-PCR
• PCR product labeling and detection kits
• PCR cloning kits
• Post-PCR purification kits 

> Combine superb polymerases and nucleotides in one easy-to-order pack
> Combine superior polymerases and nucleotides - in one convenient package
> Ensure optimal PCR performance using Roche PCR-grade nucleotides

Combine first-class polymerases and nucleotides in one easy-to-order pack

Roche Applied Science's convenient dNTPacks combine PCR-grade nucleotides, thermostable enzymes and enzyme blends, and all associated components, including specially formulated buffers with PCR-enhancing additives.

• Benefit from the attractive price
  Select proven thermostable DNA polymerases for your application. Obtain premixed solutions of PCR-Grade Nucleotides in one cost-effective package.
• Ensure superb performance
  Maximize sensitivity and performance of your amplification reactions using state-of-the-art enzymes, combined with a mixture of ultrapure PCR-Grade Nucleotides and optimized buffer components.
• Safeguard your valuable PCR reaction samples
  Conserve your time and resources using function-tested nucleotides lacking measurable contaminants.

Roche PCR-Grade Nucleotides are function-tested using RT-PCR, for proven high performance of all components. Each Roche dNTPack contains additive-free sodium salt nucleotides in a ready-to-use mix of all four nucleotides at 10 mM for each dNTP.
 

 

Combine super polymerases and nucleotides - in one convenient package

Roche Applied Science’s dNTPacks combine PCR-Grade Nucleotides, thermostable enzymes and enzyme blends, and all associated components such as buffers and PCR-enhancing additives in one convenient pack. Our highly purified nucleotides are assayed for function in RT-PCR, ensuring optimal performance of all components. Each dNTPack contains the additive-free sodium salt nucleotides as a ready-to-use mix of all 4 nucleotides (10 mM of each dNTP).
 

Click the links below for detailed information about Roche's PCR Enzyme dNTPacks:

> FastStart Taq DNA Polymerase, dNTPack  

> FastStart High Fidelity PCR System, dNTPack

> Pwo SuperYield DNA Polymerase, dNTPack

> Expand Long Range dNTPack

> Expand High Fidelity PCR System, dNTPack

> Expand High Fidelity^PLUS PCR System, dNTPack

> Expand 20 kb^PLUS PCR System, dNTPack

> GC-RICH PCR System, dNTPack

> Taq DNA Polymerase (1 U/µl), dNTPack

> Taq DNA Polymerase, dNTPack


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Ensure optimal PCR performance using Roche PCR-grade nucleotides
 

Achieve your amplification potential using Roche purified, function-tested deoxynucleotides. Conveniently use PCR-grade nucleotides available as ready-to-use mixes, comprising premixed solutions of all four nucleotides, nucleotide sets with each nucleotide in a separate vial, as well as individual nucleotides.

• Use function-tested nucleotides.
  Achieve high level PCR sensitivity (Figure 1). Each lot is tested for the absence of RNases, DNases, nicking activity and other impurities, ensuring template integrity. 
   
• Ensure consistent PCR performance.
  Choose nucleotides with a guaranteed purity of >99% dNTP and <0.9% dNDP, as determined using HPLC (Figure 2).  
   
• Achieve high stability.
  Roche PCR-Grade Nucleotides are stable for at least 30 months when stored at -15 to -25°C (Figure 3).
   

Select Roche thermostable DNA polymerases for your applications, and with Roche premixed PCR-Grade Nucleotides in economical dNTPacks.

 

Figure 1: RT-PCR of a 1849-bp fragment of the human dystrophin gene. Varying amounts of human skeletal muscle total RNA were used in this RT-PCR. Result: Roche's dNTPs produced the RT-PCR product from only 10 picograms of RNA, compared with 100 picograms when using Supplier A's dNTPs (10-fold higher sensitivity with Roche's dNTPs). Figure 3: Long term stability of dATP at -20°C. Result: Even after storage for 42 months at -20°C, Roche’s PCR-grade dATP contains 99.7% of the nucleoside triphosphate.

Figure 2: HPLC (RP 18) purity analysis of Roche's PCR-Grade Nucleotides in comparison with Supplier A's nucleotides. Result: Roche's PCR-Grade Nucleotides are free of tetraphosphates and show much higher purity overall in comparison to Supplier A's nucleotides.

Application Hints and Troubleshooting

Whom do you ask if you have trouble in the lab with the recent research? Roche Applied Science researchers stand ready to provide assistance and answers. Learn more in our "Tips from the Developer"
 

This part of our website is intended to help those who are new to PCR to get started. It will also help anyone who wants to learn ways to enhance the basic PCR technique, for instance, to increase PCR yield and fidelity or to troubleshoot problems. Because of its length, the content has been broken up into several sections.

General Considerations

• Setting up the laboratory to avoid contamination
• Special equipment
• PCR reaction components
• Thermal cycling profile for standard PCR

Factors to Consider in RT-PCR

• Choice of RT-PCR enzymes
• Choice of reverse transcription primers
• Preparation of template RNA
• Primer design
• RT-PCR procedures 
   

Prevention of Carry-over Contamination

• Preventing carry-over contamination with uracil-DNA glycosylase
• Preventing carry-over contamination in RT-PCR

Troubleshooting

• Interactive guide for solving common PCR-problems

Related Products for Conventional PCR
 

PCR is only one part of a sequence of working steps that usually begins with the preparation of a nucleic acid (DNA, RNA), sometimes followed by reverse transcription before the PCR takes place. After the PCR many different working steps can follow depending on your application, like post-PCR purification, cloning, labeling or analyzing DNA.  Roche Applied Science not only provides outstanding products for the PCR step, but also offers many reagents, kits, and other tools that can greatly facilitate your work.

• PCR-grade nucleotides
• Related products
• Mastermixes for Real-time PCR
• Products for one step RT-PCR
• Products for two step RT-PCR
• PCR product labeling kits
• Kits to detect PCR products
• PCR cloning kits
• Post-PCR purification kits

AptaTaq Fast DNA Polymerase & AptaTaq Fast PCR Master

Speed up your PCR, improve performance, and achieve results comparable to standard reactions in a fraction of time. Perform fast, convenient PCR on any thermal block cycler with two ready-to-use hot start PCR reagents:
 

• AptaTaq Fast PCR DNA Polymerase
• AptaTaq Fast PCR Master

Save Time Complete end-point PCR reactions faster with a highly processive enzyme that requires only 4 minutes of total extension time (see Figure 1). Activate the enzyme instantly through novel aptamer hotstart technology. Amplify with confidence and increase convenience Use the same enzyme and a one-for-all protocol for successful amplification of a broad variety of fragments, including GC-rich templates (see Figure 2). Set up PCR reactions at room temperature, not on ice.

 


Figure 1: Perform PCR faster than whith other supplier's "fast" PCR enzymes.
The AptaTaq Fast enzyme amplifies a 195 bp fragment from the human Erythropoietin with only 4 minutes of total extension time through 35 cycles.




Figure 2: Amplify a broad range of fragments, with at least 66% GC content.




Figure 3. Speed up RT-PCR. Combine AptaTaq Fast products with the Transcriptor Universal cDNA Master to perform convenient and spedious two step RT-PCR. Transcribe your RNA template in only 25 minutes processing time. Combine with the AptaTaq Fast PCR Master or AptaTaq Fast DNA Polymerase for spedious amplification with the one for all protocol.



Figure 4: Innovative Aptamer Technology.
AptaTaq Fast PCR Polymerase is a reversible, temperature-dependent hot start DNA amplification system with immediate activation. Similar to antibody-based methods, the enzyme is activated by heating.
Dropping the temperature below +55°C acts like a switch, shutting off polymerase activity. At temperatures above +60°C, aptaTaq Polymerase is activated, ideal for specific priming and fast PCR.