PCR and real-time PCR are sensitive and specific technologies for workflows requiring fast turnaround time and a simple workflow. Let us help you select the right reagents and Applied Biosystems instrument for your application.
Sample--preparation
  • Dynabeads magnetic beads and buffers
  • MagMAX kits
  • KingFisher Instruments
Reverse-transcription
  • Reverse transcriptases
  • One-step master mixes
  • dNTPs
Amplification
  • Master mixes
  • Polymerases
  • dNTPs
  • Primers and probes
  • Oligos
  • Plates, tubes, seals
  • Fluorescent dyes
Run-and-analysis
  • Thermal cyclers
  • Real-time PCR instruments

If your starting material is RNA—whether you’re doing gene expression analysis, viral detection applications, or any application starting with RNA, our suite of high-quality reverse transcriptase enzymes and master mixes can help you get the results you need. Many of our enzymes are also available in lyophilization-compatible (lyo-ready) formulations with less than 0.05% glycerol.

Our proprietary in vitro protein evolution technology has enabled introduction of multiple favorable mutations such as enhanced thermostability, inhibitor resistance, and processivity into the traditional reverse transcriptase.  Here are a few of our more commonly used reverse transcription enzymes.

Reverse transcriptase selection chart

Characteristic SuperScript IV Reverse Transcriptase Maxima Reverse Transcriptase RevertAid Reverse Transcriptase (M-MuLV)
Optimal reaction temperature 50°C 50°C
42°C
RNase H activity No Yes

Yes

RNase H minus version available NA Maxima H Minus RT

RevertAid H Minus RT

Reaction time 10 min 30 min 60 min
Inhibitor resistance ++++ +++

+

Lyo-ready Yes Yes Yes

Lyo-ready is a lyophilization-compatible enzyme composition without glycerol.

+= poor; ++= medium; +++= good;+++++= recommended choice.

A15299

1-step master mix

The Applied Biosystems TaqPath 1-Step RT-qPCR Master Mix offers the convenience of reverse transcription and qPCR amplification all in one reaction. It is a general-purpose reagent (GPR), and it offers enhanced inhibitor resistance and reproducibility compared to other master mixes. This GPR is manufactured in an ISO 13485 facility with measures such as change control, document control, and purchasing controls. In addition, each batch is quality control–tested prior to release.

If your starting material is DNA or cDNA—choose from our Taq DNA polymerases, or our genotyping or 2-step gene-expression master mix. Many of these products are also available as glycerol-free, lyo-ready formulations. You can choose from our most popular Taq DNA polymerases from the menu of selected enzymes below, based on a combination of hot-start technology, speed requirements, level of resistance to PCR inhibitors, or lyophilization compatibility. Our enzymes enable a high degree of sensitivity, accuracy, and reproducibility in your assays. If you require an extra level of confidence that your positives are true positives, we can also produce DNA-free enzymes by single-use technology.

Taq DNA polymerase selection chart

  Platinum II Taq Platinum Taq Amplitaq Gold LibertyTaq Wild-type Taq
Hot-start PCR Antibody-based Antibody-based Chemically modified Proprietary No
Compatible with TaqMan probes Yes Yes Yes Yes Yes
Reactivation time 2 min 2 min 10 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
Sensitivity ++++ +++ +++ + +
Sensitivity ++++ +++ +++ + +
Inhibitor resistance ++++ + + + +
Lyo-ready Yes Yes On request Yes Yes
DNA-free On request Yes On request On request On request

Lyo-ready is a lyophilization-compatible enzyme composition without glycerol.

+= poor; ++= medium; +++= good;+++++= recommended choice.

A30865

Master mixes

The TaqPath master mixes provide turnkey solutions for real-time PCR. They contain buffer, dNTPs, passive reference dye, thermostable hot-start DNA polymerase, and other components formulated for reliable 5ʹ nuclease probe-based real-time PCR. Just add your sample and Applied Biosystems TaqMan Assay components, and start your reactions.

For genotyping and copy number variation applications, our Applied Biosystems TaqPath ProAmp Master Mix offers accurate genotyping calls, even in the presence of inhibitors. It is also available in formulations with Applied Biosystems ROX or Mustang Purple dye, making it ideal for multiplexing.

For DNA detection or 2-step gene expression analysis, Applied Biosystems TaqPath qPCR Master Mix provides reproducible linear results over a wide dynamic range.

Our dNTP formulations are designed for convenience and flexibility. Standard nucleotides (dATP, dCTP, dGTP, dTTP, and dUTP) are supplied as 100 mM solutions, and nucleotide mixes can be formulated with up to 100 mM concentration for each nucleotide in the mix.

TaqMan real-time PCR assays contain optimized primer and probe sets in one convenient tube. We provide a complete range of custom-synthesized oligonucleotide primers and probes, all built to your specifications. Simply identify your target of interest or exact sequence, and our team can design your assay. In addition to custom oligonucleotides, we also offer predesigned primer and probe sets for TaqMan Assays, developed by our in-house bioinformatics team.

The TaqMan probes include a minor groove binding (MGB) moiety at the 3ʹ end that increases the melting temperature (Tm) of the probe and stabilizes probe–target hybrids. The MGB probes can therefore be significantly shorter than traditional probes, providing flexibility in designing probes for molecular test development.

The TaqMan MGB probes incorporate a non-fluorescent quencher (NFQ) to absorb (quench) the signal from the fluorescent dye label at the other end of the probe. The properties of the NFQ combined with the short length of the MGB probe result in lower background signal than with non-MGB NFQ probes. Lower background signal means increased sensitivity and precision in your data.

DNaseI

Thermo Scientific DNase I — an endonuclease that digests single- and double-stranded DNA in preparation for RT-PCR. It hydrolyzes phosphodiester bonds, producing mono- and oligo- deoxyribonucleotides with 5´-phosphate and 3´-hydroxyl groups.

Thermo Scientific dsDNase — an engineered shrimp DNase designed for rapid and safe removal of contaminating genomic DNA from RNA samples. It is an endonuclease that cleaves phosphodiester bonds in DNA to yield oligonucleotides with 5ʹ-phosphate and 3ʹ-hydroxyl termini. Its highly specific activity toward double-stranded DNA ensures that RNA and single-stranded DNA products such as cDNA and primers are not cleaved. dsDNase is easily inactivated by moderate heat treatment (55°C). These features make dsDNase an excellent choice for gDNA elimination prior to reverse transcription. It allows for a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into a one-tube procedure.

RNaseOUT

Invitrogen RNaseOUT Recombinant Ribonuclease Inhibitor — a potent noncompetitive inhibitor of pancreatic-type ribonucleases such as RNase A, and is used to avoid RNA degradation in a variety of applications. RNaseOUT Recombinant Ribonuclease Inhibitor is an acidic protein with a molecular weight of ~52 kDa. RNaseOUT reagent inhibits RNase A, RNase B, and RNase C.

Thermo Scientific Uracil-DNA Glycosylase (UDG, UNG) — prevents contamination from carryover of PCR product. The enzyme catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar, leaving an apyrimidinic site in uracil-containing single- or double-stranded DNA. The enzyme shows no activity on RNA.

When you validate your assay on Applied Biosystems real-time PCR instruments, you can leverage the expansive, global installed base in end-user laboratories. You can also take advantage of our instrument placement and reagent rental programs, which enable you to provide our instruments to your customers who don’t have access to an instrument or who need an additional instrument to ramp up capacity. As an option, you can even private-label our instruments: integrate an Applied Biosystems instrument into your workflow and deliver it to your customers with your own label attached. Your customers will also gain access to our validation and IQ/OQ (Installation Qualification/Operational Qualification) services at no additional cost. We offer a broad portfolio of high-quality qPCR instruments. For clinical applications, we recommend the Applied Biosystems QuantStudio 5, QuantStudio 7, and 7500 Fast instruments, which are available for research use only (RUO) or, in some regions, CE-IVD–cleared.

  QuantStudio 5 system QuantStudio 7 system 7500 Fast system
 
Description Modern, interactive, affordable, dual-mode A high-throughput diagnostic platform that can grow with you

A widely used workhorse
instrument

IVD instrument available? CE-IVD only Available in most regions

Available in most regions

Footprint (W x D x H) 27 x 50 x 40 cm 53 x 70 x 75 cm 34 x 45 x 49 cm
Optical system  Bright white LED White halogen lamp

White halogen lamp

Blocks Fixed block:
96-well 0.2 mL
Interchangeable:
96-well 0.2 mL
96-well 0.1 mL
384-well TaqMan Array Card
Fixed block: 96-well 0.1 mL

For Research Use Only. Not for use in diagnostic procedures.

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