Enhanced virus detection for commonly used sample types

Applied Biosystems TaqMan Fast Virus 1-Step Master Mix is a one-tube, one-step, 4X master mix designed for reliable, high-sensitivity real-time RT-PCR even in the presence of common reaction inhibitors. Its features have been selected to enhance virus detection in the kinds of research samples labs are likely to encounter.

 

High sensitivity allows viral detection in samples with low levels of target, while buffer components are formulated to handle common RT-PCR inhibitors in research samples contaminated with blood, dirt, or tissues. The mix has been optimized to work with multiple target amplicons. A single-tube format allows for uniform handling and processing for both RNA and DNA viruses while two formulation options (with and without ROX dye) enable ultimate multiplexing flexibility.


Features

Key features of TaqMan Fast Virus 1-Step Master Mixes are designed to boost virus detection:

High sensitivity

Higher-concentration master mixes allow more accurate quantification with low-titer samples or smaller reactions and fast cycling protocols with normal-titer samples.

Target flexibility

Single-run profile compatible with RNA and DNA and on any supported instrument allows for easy mix-and-match of targets on a plate.

Consistent results

Formulated to handle common RT-PCR inhibitors found in blood, stool, and other difficult research samples

Optimized for multiple targets

Excellent performance for both singleplex and multiplex reactions, with exogenous or endogenous internal controls. The formulation without ROX reference dye enables even greater flexibility for multiplexing of four targets in a single reaction well.

High efficiency

Increased qRT-PCR speed on both fast and standard instruments, speeding up your time to results and maximizing the usage of your real-time PCR instruments.


Sample data

Multiplexing capability

Because virus research often includes multiplexed primers and probes and internal reaction controls, we have optimized master mixes to work with multiple targets. TaqMan Fast Virus 1-Step master mixes demonstrate exceptional sensitivity in commonly used duplex reactions. For higher-order multiplexing, TaqMan Fast Virus 1-Step Multiplex Master Mix (No ROX) has increased flexibility to accommodate four targets in a single reaction well. By removing the ROX passive reference dye, this formulation of the master mix enables customers to use other dyes, such as JUN™ dye, in the channel.

TaqMan Fast Virus 1-Step Master Mix sensitivity in a duplex reaction

Real-time PCR reactions were performed with a viral target and an exogenous internal positive control (IPC). The virus concentration was titrated over a constant concentration of IPC. (A) TaqMan Fast Virus 1-Step Master Mix shows better sensitivity in the duplex reaction than (B) another vendor’s one-step kit—the final dilution was detectable in the linear range, rather than being undetectable. Additionally, the reaction shows superior stability of the IPC Ct values over the full dynamic range of the viral nucleic acid titration.

TaqMan Fast Virus 1-Step Multiplex Master Mix (No ROX) for higher-order multiplex reactions

A) Linear fit of ACTB, GAPDH, RNase P gene, and TMX1 targets for a 4-plex reaction with universal human reference RNA, TaqMan Fast Virus 1-Step Multiplex Master Mix (No ROX), and TaqMan Gene Expression Assays. R² = 1.0 for all targets. Amplification is observed over 6 logarithmic units for GAPDH and the RNase P gene. Amplification is observed over 4 logarithmic units for challenging targets (ACTB and TMX1). Amplification curves for (B) ACTB, (C) TMX1, (D) GAPDH, and (E) the RNase P gene.

Perform RNA and DNA virus queries next to each other

It is common for labs to test for both RNA and DNA viruses in a variety of samples. To simplify your experiments, a single TaqMan Fast Virus 1-Step Master Mix protocol has been developed to assay both types of nucleic acids, so you can perform RNA and DNA virus queries next to each other on the same plate using the same handling steps.

TaqMan Fast Virus 1-Step Master Mix in triplex

Two viral targets and an exogenous internal positive control (IPC) were run together in the same well. One target was an RNA virus, the other was a DNA virus, and the IPC was an RNA target. With a single mix optimized to both RNA and DNA viral targets in singleplex or multiplex, it is possible to maximize the efficiency of your real-time PCR instrument.

Superior inhibitor tolerance

Research samples commonly assayed for viruses include blood, dirt, and tissues. Buffer components and proprietary additives in TaqMan Fast Virus 1-Step Master Mix have been optimized to handle RT-PCR inhibitors to help ensure consistent performance even with these difficult samples, so you can be more confident in your results.

Comparison of inhibitor tolerance of TaqMan Fast Virus 1-Step Master Mix and one-step kits made by other vendors.

Three common inhibitors of RT-PCR (A, humic acid; B, hematin; C, heparin) were added to real-time RT-PCR reactions at three different concentrations with a viral target to assess the magnitude of Ct shift caused by these inhibitors. Graphs show the change in Ct from a baseline value with no inhibitor present. For all three inhibitors at all three concentrations, TaqMan Fast Virus 1-Step Master Mix results were far less affected by the inhibitor than the two competitor mixes.


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