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  • Next-generation enzymes with improved properties derived from in vitro protein evolution
  • Custom functional testing, tailored packaging, and filling to meet your assay requirements and specifications
  • Availability of lyo-ready enzyme formulations without glycerol, for compatibility with lyophilized assays
  • Manufactured in ISO 13485 or ISO 9001 certified facilities–high product quality and lot-to-lot reproducibility

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Contact us: If you would like to discuss sample availability, please contact us at MDxenzymes@thermofisher.com

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PCR Enzymes and Reverse Transcriptases

In the development of nucleic acid-based assays, the choice of PCR enzyme is key to success. Our knowledge of and expertise in enzymology and in vitro evolution allows us to develop and offer a broad portfolio of DNA polymerases based on a combination of hot-start technology, speed, level of tolerance to PCR inhibitors, and/or lyophilization compatibility (Table 1).

Benefits

Table 1. DNA polymerase selection chart.

CharacteristicsPlatinum II Taq Hot-Start DNA PolymerasePlatinum Taq DNA PolymeraseAmpliTaq Gold DNA PolymeraseLibertyTaq DNA
Polymerase
Wild type Taq DNA PolymerasePhire Hot Start II DNA Polymerase
Hot-start PCRAntibody-basedAntibody-basedChemically 
modified 
ProprietaryNo Affibody-based
TaqMan probe–compatibleYes Yes Yes Yes Yes No
Reactivation time2 min2 min10 min0 min0 min0 min
Extension rate15 sec/kb30–60 sec/kb30–60 sec/kb30–60 sec/kb30–60 sec/kb10–15 sec/kb
Sensitivity++++++++++++
Specificity+++++++++++++
Inhibitor tolerance++++++++++
Lyo-ready*On requestYesOn requestYesYesYes

* Lyo-ready is a lyophilization-compatible enzyme composition without glycerol.
Note: “+” = poor; “++” = medium; “+++” = recommended choice.

With the introduction of Thermo Scientific Phusion High-Fidelity DNA Polymerases in 2003, a new standard of performance was established for high-fidelity PCR for even the most difficult templates. Choose from our collection of high-fidelity enzymes in a variety of formats, buffers, and dNTP solutions, depending on the sophistication of your nucleic acid–based assay and your experimental requirements (Table 2).

Benefits

  • Highest fidelity on the market (>100x that of Taq polymerase)
  • Exceptional tolerance of common inhibitors of PCR 
  • Shorter cycling times for faster sample-to-results
  • Robust amplification of broad range of targets

Table 2. High-fidelity DNA polymerase selection chart.

CharacteristicsPlatinum SuperFi II DNA PolymerasePhusion Hot Start II High-Fidelity DNA PolymerasePhusion U Hot Start DNA PolymerasePhusion High-Fidelity DNA Polymerase
Fidelity vs. Taq polymerase>300x52x25x52x
Hot-start PCRAntibody-basedAffibody-basedAffibody-basedNo
Target length≤20 kb≤20 kb≤20 kb≤20 kb
Extension rate15–30 sec/kb15–30 sec/kb15–30 sec/kb15–30 sec/kb
TaqMan probe–compatibleNoNoNoNo
Inhibitor tolerance+++++++++++
dUTP toleranceNo NoYesNo
MultiplexingYesYesYesNo
Lyo-ready*On requestYesOn requestOn request

* Lyo-ready is a lyophilization-compatible enzyme composition with low glycerol (<1%).
Note: “++” = medium; “+++” = recommended choice; “++++” = outstanding.

We offer a comprehensive portfolio of RTs, from the wild type Moloney murine leukemia virus (M-MuLV) RT to the Invitrogen SuperScript line of RTs with superior characteristics such as enhanced sensitivity and reduced reaction time (Table 3). Our proprietary in vitro protein evolution technology has enabled the introduction of multiple favorable mutations that dramatically improved enzyme thermostability, resistance to inhibitors, and processivity. Our RT enzymes are also available in lyo-ready formulations, offering additional flexibility for RT-qPCR– based assay development.

Benefits

  • Robust cDNA synthesis with challenging samples
  • High efficiency and sensitivity, even in the presence of RT inhibitors
  • Enhanced thermostability and processivity
  • Minimal false-positive results with low residual host-cell DNA

Table 3. Reverse transcription selection chart.

CharacteristicsSuperScript IV RTSuperScript III RTMaxima RTRevertAid RT
(M-MuLV)
Optimal reaction temperature 50˚C50˚C50˚C42˚C
RNase H activityNoNoYesYes
RNase H minus version availableNANAMaxima H Minus RTRevertAid H Minus RT
Reaction time10 min50 min30 min60 min
Inhibitor resistance+++++++++
Sensitivity++++++++++++
Lyo-ready*YesYesYesYes

* Lyo-ready is a lyophilization-compatible enzyme composition without glycerol.
Note: “+” = poor; “++” = medium; “+++” = good; “++++” = recommended choice.

Our dNTPs have been extensively tested and verified for use in a wide variety of molecular biology applications, including highly sensitive techniques such as RT-qPCR and next-generation sequencing (Table 4).

Benefits

  • Manufactured using dedicated equipment for each dNTP
  • Tested to be free of contaminating RNase or DNase activities and inhibitors of qPCR, PCR, and RT
  • Stable for up to 36 months; dNTP preparations remain stable after >100 freeze/thaw cycles
  • High purity (≥99%) according to HPLC
  • Large-scale manufacturing (>1,000 L) available for individual dNTP solutions or mixes

Table 4. Quality testing is tailored to help ensure high performance of dNTPs in your DNA- and RNA-based assays.

Parameter*Method used
AppearanceClear, colorless solution
pH value7.3–7.5
Concentration100 ±3 mM
Base purity (HPLC)>99.5% deoxynucleoside
Purity (HPLC)≥99% triphosphate
Pyrophosphate<0.003 pmol PPi/pmol dNTP
Endodeoxyribonuclease and nicking activity Undetectable after incubation of supercoiled plasmid DNA with dNTP
Endo and exodeoxyribonucleases Undetectable after incubation of radiolabeled, single- and double-stranded oligonucleotides with dNTP
RibonucleasesUndetectable after incubation of RNA transcript with dNTP
Human DNAUndetectable in quantitative PCR test, which uses amplification of Alu repeats inhuman gDNA
E. coli DNAUndetectable in quantitative PCR test, which uses amplification of an E. coli 23S rRNA gene fragment
Functional testFunctionally tested in two-step RT-qPCR using different starting amounts of RNA transcript in reverse transcription reactions followed by amplification with hot-start Taq DNA polymerase

* Scope of standard quality-control program may vary for custom products.

PCR thermal cyclers—Rely on Applied Biosystems thermal cyclers, renowned for their dependability, accuracy, and proven performance since 1987. Our thermal cyclers can be customized to fit your platform, including software and private label.

PCR plastics—Take advantage of precision manufacturing with high-grade plastic consumables.

 Download the OEM PCR and qPCR Plastics Guide

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