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SYBR™ Green PCR Master Mix

The SYBR® Green PCR Master Mix, optimized for real-time PCR analysis, conveniently combines SYBR Green 1 Dye, AmpliTaq Gold® DNA Polymerase, dNTPs with dUTP, Passive Reference 1, and optimized buffer components in easy-to-use premix vials.


• Easy-to-use solution contains all components needed for PCR applications using SYBR® Green dye, which minimizes assay set-up time
• Used with rapid assay development guidelines, that minimize optimization time
• SYBR® Green I dye detects double-stranded DNA, so specific probes are not required
• Includes Passive Reference I, which is required for signal normalization in all SYBR® Green dye reactions
• Contains AmpliTaq Gold® DNA polymerase, which allows the highest performance available by minimizing non-specific product formation (including primer-dimer)
• Optimized buffer components are proprietary buffer enhancements that add to the performance and reliability of the assay using SYBR® Green dye
• Recommended storage conditions are 2°-8°C, so no thawing is required before use, thereby minimizing assay set-up time


Applied Biosystems has developed a comprehensive set of guidelines to ensure success when using sequence detection systems (SDS) reagents and instrumentation. These guidelines are simple and easy to follow. Many traditional variables, such as magnesium chloride concentration and the thermal cycling protocol itself, have been standardized, greatly reducing assay development time.

All SDS reagent kits contain a passive internal reference to normalize non-PCR-related fluorescence fluctuations. Normalizing with a passive internal reference minimizes well-to-well variability that result from a variety of causes, such as pipetting errors and sample evaporation.

Real-time quantitative PCR was first performed using the 5' fluorogenic nuclease assay, which uses a TaqMan® probe during the PCR amplification to provide added specificity. An additional chemistry is also available, using SYBR® Green I dye, that can provide real-time quantitative PCR information. Assays using the SYBR® Green 1 double-stranded DNA binding dye do not require a TaqMan® probe and provide additional experimental flexibility.

The incorporation of SYBR Green I dye into a real-time PCR reaction lets the user detect any double-stranded DNA generated during PCR. This provides great flexibility because no target specific probes are required, and yet both specific and non-specific products will generate signal. The use of the hot-start enzyme AmpliTaq Gold® DNA Polymerase in all SYBR Green I reagent kits yields the highest performance available by minimizing non-specific product formation. SYBR Green I dyes are ideal for use in target identification (screening assays), or when only a small number of reactions are required for a given assay.

Alternative product
Try PowerUp SYBR Green Master Mix, our newest, high-performance, SYBR dye-based master mix for superior performance at a very competitive price. With PowerUp SYBR Green Master Mix, we’ve taken the best of SYBR Green PCR Master Mix and added additional capabilities for your gene expression analysis.

TaqMan™ Fast Advanced Master Mix (Applied Biosystems™)

TaqMan Fast Advanced Master Mix delivers accurate quantification and dependability in less time on an array of qPCR instrument platforms. It provides best in class performance, including the widest quantifiable range, in both single and duplex reactions, even with challenging targets. TaqMan Fast Advanced Master Mix contains AmpliTaq Fast DNA Polymerase, uracil-N-glycosylase (UNG), dNTPs with dUTP, ROX dye (passive reference), and optimized buffer components. It is supplied at a 2X concentration.

Features of the TaqMan Fast Advanced Master Mix include:
Best-in-class performance—superior sensitivity, accuracy, dynamic range, and specificity compared to standard mixes in standard mode
Engineered for enhanced benchtop stability—stable at room temperature for up to 72 hours in preassembled reactions
Optimized for multiplexing—validated for duplexing with exogenous and endogenous internal positive control assays
Reduced run times (<40 minutes) on fast and standard instrumentation—optimized for fast mode on fast instruments and fast cycling conditions on standard instruments
Seamlessly transitions into your experiments—validated with TaqMan assays for gene expression and microRNAs, and TaqMan array micro-fluidic cards

Best-in-class performance
TaqMan Fast Advanced Master Mix has been designed to provide performance equal to or better than standard master mixes. It has been benchmarked against the leading suppliers' standard and fast master mixes to help ensure that it succeeds in providing best-in-class sensitivity, accuracy, dynamic range, and specificity. TaqMan Fast Advanced Master Mix’s dynamic range (up to 7 logs) is the widest in the industry. The impressive sensitivity of the mix is showcased in the figure below which compares the Ct values of TaqMan Fast Advanced Master Mix and TaqMan Universal PCR Master Mix across a panel of gene expression assays.

Benchtop stability for high-throughput handling and convenience
TaqMan Fast Advanced Master Mix has been engineered to retain a high level of performance in preassembled reactions for up to 72 hours. The stability of this mix provides users of high-throughput liquid handling systems the assurance that the results on the first plate will mimic those of the last plate. The figure below shows an assay that was run upon assembly (time 0) and after 72 hours of incubation at 30°C, simulating the most extreme room temperature scenario.

For those with less extreme throughput needs, the enhanced stability of this master mix provides an overall added convenience to your workflow, as you are no longer constrained to immediately running plates upon assembly.

Optimized for multiplexing
We realize that confidence is paramount when it comes to your results. For added confidence in the results in every well, TaqMan Fast Advanced Master Mix has been designed to help deliver accurate results for duplex reactions using an internal positive control (IPC). The figure below shows results for the experimental target β-actin (gene name: ACTB), which was serially diluted and amplified in single-target reactions and duplex reactions.

Validated for microRNA assays
TaqMan Fast Advanced Master Mix has been validated for use with multiple real-time PCR applications, including microRNA assays. The formulation provides high specificity and dynamic range, the two most critical performance attributes that define successful results when working with microRNAs. The data in the figure below demonstrate excellent PCR linearity over a 6-log range of input template.

Reduced run times on standard instrumentation
TaqMan Fast Advanced Master Mix has been optimized for use with both fast and standard instrumentation, enabling researchers who currently own standard instruments to also reap the performance benefits and time savings this mix provides. The figure below showcases the impressive results achieved when using TaqMan Fast Advanced Master Mix under fast thermal cycling conditions on the Applied Biosystems 7300 Real-Time PCR System. The mix has been tested with all Applied Biosystems standard real-time PCR instrumentation (7900HT, 7500, and 7300 systems) to enable success independent of whether or not you own a fast-enabled instrument.

TaqMan™ PreAmp Master Mix (Applied Biosystems™)

TaqMan® PreAmp Master Mix pre-amplifies small amounts of cDNA without introducing amplification bias to the sample. Preamplification enables you to stretch your precious sample into many more real-time PCR reactions.

TaqMan® PreAmp Master Mix lets you:

• Analyze mRNA from precious samples such as laser capture microdissections, needle biopsies, and formalin-fixed paraffin-embedded (FFPE) tissues
• Stretch as little as 1 ng of cDNA into 200 real-time PCR reactions for gene expression analysis using TaqMan® Gene Expression assays
• Analyze up to 100 gene expression targets with minimal hands-on time

Preserves Equilibrium of Targets Without Preamplification Bias
In the past, preamplification kits and techniques often resulted in uneven amplification of targets and incorrect or biased data (as shown by low correlation coefficients between preamplified and unamplified samples). TaqMan® PreAmp Master Mix amplifies with no bias and provides extremely high correlation between amplified and unamplified cDNA.

Easy Workflow
With TaqMan® PreAmp Master Mix, preamplification setup takes only a few minutes. After a one-time pooling of the TaqMan® assays , preamplification setup (mixing the pooled assays with cDNA sample and TaqMan® PreAmp Master Mix) takes only five additional minutes. Cycling takes 1.5 hours using a GeneAmp® 9700 thermal cycler.

QuantStudio™ 12K Flex OpenArray™ Gene Expression Starter Kit (Applied Biosystems™)

A variety of starter kits are available to introduce you to the various applications of the QuantStudio™12K Flex OpenArray® sample block. Each starter kit contains 2 arrays, plus sufficient reagents and accessories to perform two experiments. The Gene Expression starter kit contains 2 gene expression endogenous control plates. The Gene Expression kit also contains cDNA controls to serve as samples for your experiments. Note that the QuantStudio™12K Flex Software (included with the QuantStudio™ 12K Flex System) offers example experiment files for each application.

The accessories provided in the kit include: 6 lids and plugs, 6 immersion fluid syringes, 2 carriers, 1 box of Accufill™ System tips, and 384-well plates.

ABsolute Blue QPCR Mix, SYBR Green, with separate ROX vial (Thermo Scientific™)

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Alternative Product: Try PowerUp SYBR Green Master Mix, our newest, high-performance, SYBR dye-based master mix for superior performance at a very competitive price. With PowerUp SYBR Green Master Mix, we’ve taken the best of ABsolute Blue QPCR Mix, SYBR Green, and added additional capabilities for your gene expression analysis.

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Thermo Scientific ABsolute Blue qPCR SYBR Green Mixes are optimized 2X master mixes for SYBR Green chemistry. The master mixes incorporate an inert blue dye to significantly enhance the contrast between reagent and plastic, making verification of master mix dispensing quick, easy, and foolproof. Absolute Blue qPCR Mixes include Thermo-Start Taq hot start enzyme for increased sensitivity.

Highlights

High performance—ABsolute sensitivity, consistency & convenience
Enhanced contrast and reduced errors—easily identify correctly aliquotted reaction mix in all plates and tubes
Universal—compatible with all major qPCR platforms
Adaptable—available with ROX or Fluorescein passive reference dyes

Applications

• Gene expression analysis
• siRNA validation
• Genotyping

Related Products
ABsolute Blue QPCR Mix, low ROX
ABsolute Blue QPCR Mix, with separate ROX vial
ABsolute Blue QPCR Mix, ROX
ABsolute Blue QPCR Mix, SYBR Green, low ROX
ABsolute Blue QPCR Mix, SYBR Green, ROX
ABsolute Blue QPCR Mix, SYBR Green, with separate ROX vial
ABsolute Blue QPCR Mix, SYBR Green, fluorescein

AgPath-ID™ One-Step RT-PCR Reagents (Applied Biosystems™)

AgPath-ID™ One-Step RT-PCR Reagents are designed for sensitive, robust amplification of RNA targets using a rapid single-tube TaqMan® real-time reverse transcription PCR (RT-PCR) strategy.
• Recommended for RNA pathogen amplification
• Consistent amplification of RNA targets with high specificity and sensitivity
• Optimized to work with your target-specific primers and probes
• Contains ROX for quantitative fluorescent signal normalization

AgPath-ID™ One-Step RT-PCR Reagents are configured for fast and simple reaction setup. The reactions are assembled in a single tube, minimizing sample handling errors and expediting set-up time. Once assembled, results are available in approximately one hour. The 25X RT-PCR Enzyme Mix included in the kit contains highly efficient ArrayScript™ Reverse Transcriptase, a mutant MMLV RT that produces high cDNA yields, and AmpliTaq Gold® polymerase, the preferred hot-start DNA polymerase for specific target amplification. The 2X RT-PCR Buffer has been optimized for efficient, robust reverse transcription and PCR includes the passive reference dye, ROX™ Dye, for quantitative fluorescent signal normalization. A Detection Enhancer is also provided as an optional reagent for amplification of templates with a high GC content or persistent secondary structure.

CellsDirect™ One-Step qRT-PCR Kit with ROX, for ABI 7000, 7300, 7700, and 7900(HT) instruments (Invitrogen™)

The CellsDirect™ One-Step qRT-PCR Kit delivers highly sensitive and specific real-time qPCR results directly from cells, without the need for an RNA purification step.

Features of the CellsDirect™ One-Step qRT-PCR Kit include:

Efficiency—Offers significant time and cost savings for increased throughput
Convenience—Enzyme mix includes both SuperScript® III Reverse Transcriptase and Platinum® Taq DNA Polymerase
Sensitivity—No separate RNA isolation step increases sensitivity by minimizing sample loss
Broad Detection Range—Detect targets from samples ranging from a single cell to 10,000 cells
Versatility—Compatible with a wide variety of cell lines and samples collected by Laser Capture Microdissection (LCM)

Streamline Your Workflow
In traditional real-time quantitative RT-PCR (qRT-PCR), you first isolate RNA from cells in a time-consuming procedure that can lead to loss of material. Using the CellsDirect™ One-Step qRT-PCR Kit, you lyse the cells and add the complete lysate directly to a one-step qRT-PCR reaction with minimal handling and sample loss.

Ideal for Your qRT-PCR Application
CellsDirect™ technology is validated for gene expression studies involving limited samples, and is suitable for validation of RNAi knockdowns. CellsDirect™ One-Step qRT-PCR Kits are especially valuable for high-throughput applications by eliminating costly, time-consuming RNA purification steps (Figure 1). The CellsDirect™ One-Step qRT-PCR Kit may also be used to detect genomic DNA targets.

CellsDirect™ One-Step qRT-PCR Kits may be used in conjunction with hydrolysis probes (e.g. TaqMan®) or LUX™ fluorogenic primers.

Note: This kit includes 2X Reaction Mix containing ROX™ Reference Dye. An equivalent kit (11753-500), containing a separate tube of ROX™ Reference Dye, is also available.

TaqMan™ Universal PCR Master Mix, no AmpErase™ UNG (Applied Biosystems™)

TaqMan Universal PCR Master Mix, No AmpErase UNG, is the ideal reagent solution when you need a master mix for multiple 5' nuclease DNA applications. Applied Biosystems reagents have been validated with TaqMan assays and Applied Biosystems real-time systems to ensure sensitive, accurate, and reliable performance every time.

• Contains AmpliTaq Gold DNA Polymerase to provide a better yield and a more robust 5' nuclease assay than AmpliTaq DNA Polymerase
• Includes a passive internal reference based on proprietary ROX dye, for superb precision on Applied Biosystems real-time PCR instruments
• Buffer enhancements guarantee performance and reliability in all applications, even with G/C-rich sequences
• Contains dUTP in the dNTP mix to protect against carry-over contamination, when used with AmpErase UNG
• Rapid assay development guidelines are provided to minimize optimization time

Alternative product: Try TaqMan Fast Advanced Master Mix, our highest-performance, probe-based master mix. With TaqMan Fast Advanced Master Mix, we’ve taken the best of TaqMan Universal PCR Master Mix and added additional capabilities for your gene expression analysis.

QuantStudio™ 12K Flex OpenArray™ Vaginal Microbiota Starter Kit (Applied Biosystems™)

A variety of starter kits are available to introduce you to the various applications of the QuantStudio™ 12K Flex OpenArray™ sample block. Each starter kit contains 2 arrays, plus sufficient reagents and accessories to perform two experiments. The QuantStudio™ 12K Flex OpenArray™ Vaginal Microbiota Starter Kit contains two training plates and a synthetic amplification control to serve as a sample for your experiments. This starter kit serves as a training tool for researchers new to this application.

The accessories provided in the kit include: 6 lids and plugs, 6 immersion fluid syringes, 2 carriers, 1 box of Accufill™ System tips, and 384-well plates.

VetMAX™ PRRSV NA & EU Reagents (Applied Biosystems™)

Note: The name of this product has changed. It was previously called PRRSV REAGENTS W/PATCH EACH.

VetMAX™ NA and EU PRRSV Reagents contains primers/probes that enable multiplex, one-step, reverse transcription and real-time PCR (RT-PCR) targeting North American and European Porcine Reproductive and Respiratory Syndrome Virus (PRSSV) RNA and Xeno™ RNA Control. VetMAX™ NA and EU PRRSV Reagents contains an updated primer/probe mix compared to the similar TaqMan® NA and EU PRRSV Reagents.

• Rapid and cost-effective method for amplification of target sequences
• Quality reagents, manufactured under ISO 9001, providing confidence in the supply and quality of products
• Over 20 years in the DNA/RNA arena, with extensive bioinformatics capabilities and experience

PRSSV causes significant losses in the swine industry due to reproductive disorders, respiratory conditions, and growth retardation. PRRSV can be detected in blood and semen 24–48 hours after infection. Real-time PCR is the most sensitive method for monitoring RNA amplification.

ABsolute Blue QPCR Mix, SYBR Green, fluorescein (Thermo Scientific™)

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Alternative Product: Try PowerUp SYBR Green Master Mix, our newest, high-performance, SYBR dye-based master mix for superior performance at a very competitive price. With PowerUp SYBR Green Master Mix, we’ve taken the best of ABsolute Blue QPCR Mix, SYBR Green, and added additional capabilities for your gene expression analysis.

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Thermo Scientific ABsolute Blue qPCR SYBR Green Mixes are optimized 2X master mixes for SYBR Green chemistry. The master mixes incorporate an inert blue dye to significantly enhance the contrast between reagent and plastic, making verification of master mix dispensing quick, easy, and foolproof. Absolute Blue qPCR Mixes include Thermo-Start Taq hot start enzyme for increased sensitivity.

Highlights

High performance—ABsolute sensitivity, consistency & convenience
Enhanced contrast and reduced errors—easily identify correctly aliquotted reaction mix in all plates and tubes
Universal—compatible with all major qPCR platforms
Adaptable—available with ROX or Fluorescein passive reference dyes

Applications

• Gene expression analysis
• siRNA validation
• Genotyping

Related Products
ABsolute Blue QPCR Mix, low ROX
ABsolute Blue QPCR Mix, with separate ROX vial
ABsolute Blue QPCR Mix, ROX
ABsolute Blue QPCR Mix, SYBR Green, low ROX
ABsolute Blue QPCR Mix, SYBR Green, ROX
ABsolute Blue QPCR Mix, SYBR Green, with separate ROX vial
ABsolute Blue QPCR Mix, SYBR Green, fluorescein

TaqPath™ 1-Step Multiplex Master Mix (No ROX) (Applied Biosystems™)

TaqPath 1-Step Multiplex Master Mix (No ROX) is a fast, single-tube, 4X RT-qPCR mix that provides for sensitive, reproducible detection of up to four different RNA/DNA targets in a single multiplex reaction. Particularly useful for diagnostics, virus detection, and high throughput gene expression workflows, the mix includes thermostable MMLV reverse transcriptase, dNTPs, UNG, and thermostable Fast DNA polymerase all in a single tube. TaqPath is manufactured under a strong ISO13485 quality management system and is designed to produce consistent product performance lot after lot.

Request an evaluation sample >

This single-tube, 4X format facilitates easy reaction set up—just add user-supplied assay and sample (Figure 1). With reproducible performance even in the presence of inhibitors, coupled with stringent quality control to minimize lot-to-lot variation, TaqPath 1-Step Multiplex Master Mix (No ROX) is a superior choice for your multiplex diagnostic testing or development needs.

Features of the TaqPath 1-Step Multiplex Master Mix include:
• High sensitivity to detect low-copy targets with reproducible CT results
• Six logs of dynamic range with both RNA and DNA targets
• Optimized for multiplexing up to four targets simultaneously without a passive reference dye
• Tolerant of inhibitors commonly found in clinical samples

High sensitivity
To enable reproducible detection of low-titer pathogen and transcripts, TaqPath 1-Step Multiplex Master Mix (No ROX) has been optimized as a higher-concentration 4X master mix that allows you to input more sample into each reaction, increasing sensitivity even in low-volume reactions. Figure 1 shows the consistency of results obtained when detecting 10 copy inputs of RNA target.

Wide dynamic range compatible with RNA and DNA
TaqPath 1-Step Multiplex Master Mix (No ROX) has been optimized to provide high specificity and dynamic range for use with both RNA and DNA targets. Since virology labs often test for both RNA and DNA viruses, TaqPath 1-Step Multiplex Master Mix (No ROX) is designed to use a single protocol to assay both types of nucleic acid. This input flexibility can help streamline the number of different workflows in your lab to improve efficiency. Figure 2 demonstrates the excellent PCR linearity across a 6-log RNA dilution series, while Figure 3 exhibits a 6-log linear dynamic range for both RNA and DNA targets.

Optimized for multiplexing
TaqPath 1-Step Multiplex Master Mix has been validated for multiplexing up to four targets simultaneously, allowing for additional targets and/or controls to be run simultaneously for efficiency or quality control purposes. The mix does not include a passive reference dye, enabling the measurement of the JUN dye, or a similar emission wavelength dye, in the channel previously used to measure ROX dye. Figure 3 demonstrates TaqPath 1-Step Multiplex Master Mix (No ROX) performance in 4-plex reactions with both RNA and cDNA targets.

Inhibitor tolerant
Unlike other master mixes on the market, the unique proprietary formulation of TaqPath 1-Step RT-qPCR Master Mix (No ROX) allows robust performance even in the presence of substances that can normally inhibit PCR, such as heparin, hematin, or EDTA, increasing your confidence when working with a variety of complex clinical samples. TaqPath master mixes demonstrate increased tolerance of inhibitors compared to competitor mixes (Figure 4).

Manufacturing production and process controls
TaqPath 1-Step Multiplex Master Mix (No ROX) is manufactured under a strong ISO13485 quality management system that uses traceable quality raw materials and validated operating procedures. With established controls from purchasing through QC release, TaqPath product manufacturing is designed to produce consistent product performance lot after lot. Figure 5 demonstrates CT lot-to-lot consistency is preserved across multiple assays with different attributes and input target levels.

Broad instrument compatibility
The TaqPath 1-Step Multiplex Master Mix (No ROX) formulation can be run in either fast or standard cycling conditions with equivalent performance across a wide variety of real-time PCR systems including the Applied Biosystems 7500, 7500 Fast, 7500 Fast Dx, QuantStudio 3, 5, 6, 7, 12k, Dx, and ViiA 7 real-time PCR systems.

General purpose reagent
TaqPath 1-Step RT-qPCR Master Mix is a General Purpose Reagent (GPR), labeled for laboratory use.

Related products
TaqPath 1-Step Master Mix is available in three different formulations:
TaqPath 1-Step qRT-PCR Master Mix—includes ROX dye as the passive reference
TaqPath 1-Step Multiplex Master Mix—includes MUSTANG PURPLE dye as the passive reference
TaqPath 1-Step Multiplex Master Mix (no ROX) (this page)—does not include a passive reference dye

Path-ID™ Multiplex One-Step Kit (Applied Biosystems™)

The Path-ID™ Multiplex One-Step RT-PCR Kit is designed for multiplex, quantitative, reverse transcription PCR (qRT-PCR). It is optimized for the amplification of up to four targets simultaneously using your RNA samples and TaqMan® primer⁄probe sets. Reactions are run using a single-tube, one-step procedure to reverse-transcribe the RNA and amplify your targets. The kit contains a Multiplex Enzyme Mix with Arrayscript™ reverse transcriptase and AmpliTaq Gold® DNA Polymerase, and a buffer mix.

ABsolute QPCR Mix, no ROX (Thermo Scientific™)

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Alternative Product: Try TaqMan Fast Advanced Master Mix, our highest-performance, probe-based master mix. With TaqMan Fast Advanced Master Mix, we’ve taken the best of ABsolute QPCR Mix and added additional capabilities for your gene expression analysis.

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Thermo Scientific ABsolute qPCR Mixes are optimized for use with hybridization probes and contain all the components necessary to perform quantitative PCR, with the exception of template and primers/probes. The 2X master mix contains a proprietary reaction buffer that provides high sensitivity, specificity, and reproducibility. ABsolute qPCR Mixes are available with the option of included high ROX reference dye concentration, Low ROX concentration, and with no ROX reference dye. There is also a master mix version optimized for capillary instruments.

Highlights

• High sensitivity & specificity - The master mixes contain our patented hot start enzyme Thermo-Start DNA Polymerase
• Reproducibility - All mixes undergo extensive QC to ensure batch to batch consistency
• Optimised buffer - For maximal performance in multiplex reactions

Applications
• Gene expression analysis
• siRNA validation
• Genotyping

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ABsolute QPCR Mix, SYBR Green, ROX
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AgPath-ID™ One-Step RT-PCR Reagents with manual (Applied Biosystems™)

AgPath-ID™ One-Step RT-PCR Reagents are designed for sensitive, robust amplification of RNA targets using a rapid single-tube TaqMan® real-time reverse transcription PCR (RT-PCR) strategy.
• Recommended for RNA pathogen amplification
• Consistent amplification of RNA targets with high specificity and sensitivity
• Optimized to work with your target-specific primers and probes
• Contains ROX for quantitative fluorescent signal normalization

AgPath-ID™ One-Step RT-PCR Reagents are configured for fast and simple reaction setup. The reactions are assembled in a single tube, minimizing sample handling errors and expediting set-up time. Once assembled, results are available in approximately one hour. The 25X RT-PCR Enzyme Mix included in the kit contains highly efficient ArrayScript™ Reverse Transcriptase, a mutant MMLV RT that produces high cDNA yields, and AmpliTaq Gold® polymerase, the preferred hot-start DNA polymerase for specific target amplification. The 2X RT-PCR Buffer has been optimized for efficient, robust reverse transcription and PCR includes the passive reference dye, ROX™ Dye, for quantitative fluorescent signal normalization. A Detection Enhancer is also provided as an optional reagent for amplification of templates with a high GC content or persistent secondary structure.