PCR Enzymes and Reverse Transcriptases for Commercial Supply
Innovation and quality to drive your ideas
- 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
If you would like to discuss sample availability, please contact us at MDxenzymes@thermofisher.com
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 resistance to PCR inhibitors, and/or lyophilization compatibility (Table 1).
- High sensitivity and specificity for detection of low-copy DNA targets
- Minimal activation time for faster sample-to-results
- Robust amplification of difficult targets, including those of suboptimal purity
- Lyophilization-compatible (lyo-ready) formulations available
Table 1. DNA polymerase selection chart.
|Characteristics||PlatinumTaq DNA Polymerase, inhibitor resistant||Platinum Taq DNA Polymerase||AmpliTaq Gold DNA Polymerase||LibertyTaq DNA
|Wild type Taq DNA polymerase||Phire Hot Start II DNA Polymerase|
|Reactivation time||2 min||2 min||10 min||0 min||0 min||0 min|
|Extension rate||15–30 sec/kb||30–60 sec/kb||30–60 sec/kb||30–60 sec/kb||30–60 sec/kb||10–15 sec/kb|
|Lyo-ready*||On request||Yes||On request||Yes||Yes||Yes|
*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).
- 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.
|Characteristics||Platinum SuperFi DNA Polymerase||Phusion Hot Start II High-Fidelity DNA Polymerase||Phusion U Hot Start DNA Polymerase||Phusion High-Fidelity DNA Polymerase|
|Fidelity vs. Taq polymerase||>100x||52x||25x||52x|
|Target length||≤20 kb||≤20 kb||≤20 kb||≤20 kb|
|Extension rate||15–30 sec/kb||15–30 sec/kb||15–30 sec/kb||15–30 sec/kb|
|Lyo-ready*||On request||Yes||On request||On 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.
- 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.
|Characteristics||SuperScript IV RT||SuperScript III RT||Maxima RT||RevertAid RT
|Optimal reaction temperature||50˚C||50˚C||50˚C||42˚C|
|RNase H activity||No||No||Yes||Yes|
|RNase H minus version available||NA||NA||Maxima H Minus RT||RevertAid H Minus RT|
|Reaction time||10 min||50 min||30 min||60 min|
* 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).
- 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.
|Appearance||Clear, colorless solution|
|Concentration||100 ±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|
|Ribonucleases||Undetectable after incubation of RNA transcript with dNTP|
|Human DNA||Undetectable in quantitative PCR test, which uses amplification of Alu repeats inhuman gDNA|
|E. coli DNA||Undetectable in quantitative PCR test, which uses amplification of an E. coli 23S rRNA gene fragment|
|Functional test||Functionally 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.
For Research Use Only. Not for use in diagnostic procedures.