Shop All qRT-PCR Kits

CellsDirect™ One-Step qRT-PCR Kit 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. CellsDirect™ One-Step qRT-PCR Kits are especially valuable for high-throughput applications by eliminating costly, time-consuming RNA purification steps (see figure). 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 a separate tube of ROX™ Reference Dye. An equivalent kit (11754-500) with ROX™ Reference Dye included in the 2X Reaction Mix is also available.

EXPRESS One-Step Superscript™ qRT-PCR Kit, universal Invitrogen™

EXPRESS One-Step SuperScript qRT-PCR kits incorporate proven Invitrogen enzymes specifically tailored to deliver superior results on high-throughput, fast-cycling instruments for your probe-based qPCR experiments. This fast, one-step mix is designed to work with a variety of instruments’ default protocols, including Applied Biosystems and non-Applied Biosystem (see universal version) qPCR platforms.

Features of the EXPRESS One-Step Superscript qRT-PCR Kit include:
Fast-activating, antibody-mediated Platinum Taq DNA Polymerase–complete, rapid activation; provides high sensitivity and reproducibility
The One-Step SuperScript III Reverse Transcriptase module– a unique formulation to ensure high yield and less PCR inhibition
Flexible formats with separate or pre-mixed ROX–designed to work with default instrument protocols on a wide range of high-throughput fast instruments (e.g., AB 7500 Fast, AB 7900 HT Fast, AB StepOne, Roche LightCycler® 480, Corbett Rotor-Gene™)
Contains heat-labile UDG–significantly reduces carryover contamination risk with excellent activity from room temperature to approximately 40°C. Due to the thermostability of SuperScript III, the cDNA synthesis step is run at 50°C, at which temperature the heat-labile UDG is totally and irreversibly inactivated

EXPRESS kits work with manufacturer’s default protocols on fast instruments and come in a convenient, stable SuperMix format for ultimate convenience. The kits are offered with either pre-mixed ROX or a separate tube of ROX to fit your instrument needs, as well as SYBR Green chemistry options:
EXPRESS One-Step Superscript qRT-PCR Kit, universal (this page)
EXPRESS One-Step Superscript qRT-PCR Kit, with premixed ROX
EXPRESS One-Step SYBR GreenER Kit, with premixed ROX
EXPRESS One-Step SYBR GreenER Kit, universal

Single Cell-to-CT™ qRT-PCR Kit Invitrogen™

The Ambion® Single Cell-to-CT™ Kit contains a complete validated workflow for gene expression analysis for samples containing 1–10 cells. Each kit contains reagents for sample preparation, reverse transcription, pre-amplification, and qPCR that have been optimized together in a simple workflow that can be completed in only 5 steps (see figure).

Key product features:

• Pre-optimized workflow for real-time RT-PCR from single cells
• Maximum sensitivity for single-cell analysis
• Superior performance and reproducibility compared to alternative methods
• Complete kit convenience and easy-to-follow protocol

Pre-optimized protocol ensures success and saves time
Each workflow begins with a simple 7-minute sample prep, where cells are effectively and reproducibly lysed with minimal processing into a solution that is compatible with downstream RT-PCR, without the need for purification. This is followed by the use of Superscript® RT for reverse transcription, the TaqMan® PreAmp Master Mix to amplify the cDNA, and TaqMan® Gene Expression Master Mix for qPCR. All four of these steps have been developed to provide maximum sensitivity. The entire cell sample is maintained in the same well throughout the procedure so there is no sample loss or dilution of the precious limited material that could affect sensitivity.

Single cell sensitivity
Sensitivity of real-time PCR results can be affected by the efficiency of sample preparation, reverse transcription, or amplification. The Single Cell-to-CT™ Kit has addressed each of these potential problem areas to create a solution that enables reliable and robust gene expression analysis from single cells with maximal sensitivity. Detection of single cell equivalents using the Single Cell-to-CT™ Kit demonstrates appropriate linearity and sensitivity of qPCR results compared to a 100 cell sample control with excellent technical reproducibility (see figure). As expected, individual single cells exhibit greater variability due to inherent biological heterogeneity. In addition, through inclusion of a cDNA pre-amplification step, targets of interest are accurately amplified prior to real-time PCR. This critical step boosts signal in an unbiased manner, extending the analysis potential of limited samples. The accuracy and signal enhancement of a 96-gene panel using pre-amplification is shown (see figure).

Superior performance and reproducibility compared to alternative methods
Traditional sample preparation methods using organic solvents, glass fiber filters, or magnetic beads are not suitable for single-cell analysis due to the loss of sample through incomplete RNA precipitation, binding, and elution. Additionally, most single-cell homebrew methods which involve simple boiling to lyse cells lack the ability to inactivate endogenous RNases to stabilize gene expression profiles and can result in chemical cleavage of the RNA and inhibitor carryover, including gDNA contamination, which can affect all qRT-PCR applications. In contrast, the Single Cell-to-CT™ Kit shows superior sensitivity and reproducibility for single-cell analysis compared to traditional purification or homebrew boiling methods (see figure).

SuperScript™ III Platinum™ SYBR™ Green One-Step qPCR Kit w/ROX Invitrogen™

Our One-Step qRT-PCR Kits with ROX combine the most powerful reverse transcriptase and DNA polymerase technologies to deliver precise and accurate analysis of gene expression in a convenient, high-throughput one-step format. One-Step qRT-PCR kits with ROX include:


–SuperScript® III Reverse Transcriptase: produces higher yields of full-length cDNA for broader gene representation

–Platinum® Taq DNA Polymerase: activates faster and more completely than other hot-start enzymes to deliver more sensitive amplification (Figure 1)
–ROX reference dye premixed at optimal concentration for ABI instruments



Two kit configurations allow your choice of detection methods:

–SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR Kit provides a highly efficient, cost-effective qRT-PCR system with SYBR Green I detection
–SuperScript® III Platinum® One-Step qRT-PCR Kit provides sensitive, specific detection for use with LUX™ fluorogenic primers or dual-labeled probes (e.g. TaqMan®) (Figure 2)


Contents and Storage:
SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR Kit with ROX includes SuperScript® III RT/Platinum® Taq Mix, 2X Reaction Mix (containing SYBR® Green I, ROX Reference Dye, dNTPs, and MgSO4), and 50 mM MgSO4. SuperScript® III Platinum® One-Step qRT-PCR Kit with ROX includes SuperScript® III RT/Platinum® Taq Mix, 2X Reaction Mix (containing ROX Reference Dye, dNTPs and MgSO4), and 50 mM MgSO4. Store all components at -20°C. Store ROX Reference dye in the dark. Guaranteed stable for 6 months when properly stored.

EXPRESS One-Step SYBR™ GreenER™ Kit, universal Invitrogen™

EXPRESS One-Step SYBR GreenER kits are compatible with fast-cycling instruments for your SYBR-based qPCR experiments. This kit comes with a separate tube of ROX reference dye. EXPRESS One-Step SYBR GreenER SuperMix kits with premixed ROX reference dye are also available.

This kit provides:
• Fast-activating, antibody-mediated Platinum™ Taq DNA Polymerase
• The One-Step SuperScript™ III Reverse Transcriptase for one-step qRT-PCR
• SYBR GreenER dye
• UDG carryover protection—uracil-DNA-glycosylase (UDG)/dUTP to reduce carryover contamination

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

Power SYBR™ Green RNA-to-CT1-Step Kit Applied Biosystems™

Get sensitivity and specificity in an easy-to-use one-step qRT-PCR reaction. The Power SYBR® Green RNA-to-CT1-Step kit has been formulated to ensure maximum sensitivity and reliability. The novel formulation reduces false positive results caused by primer dimers, significantly improves data accuracy, and provides reliable detection of low abundance targets. It includes:
• AmpliTaq® Gold DNA Polymerase UP for hot start qPCR, minimizing non-specific product formation
• ArrayScript™ UP Reverse Transcriptase, an engineered RT that produces high yields of full-length cDNA
• An additive that reduces primer dimer formation, reducing non-specific products that can cause false positive signals
• RNase inhibitor, to prevent degradation of RNA templates
• A proprietary ROX™ dye that serves as a passive internal reference to normalize non-PCR-related fluorescence fluctuations for superb precision on Applied Biosystems® real-time PCR instruments.

SuperScript™ III Platinum™ SYBR™ Green One-Step qRT-PCR Kit Invitrogen™

The SuperScript® III Platinum® One-Step qRT-PCR Kits combine the most powerful reverse transcriptase and DNA polymerase technologies to deliver precise and accurate analysis of gene expression in a convenient one-step format. The SuperScript® III Platinum® One-Step qRT-PCR Kit provides sensitive, specific detection with fluorogenic primers and probes, while the SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR Kit offers the ease and convenience of SYBR® Green I detection

SuperScript® III Platinum® One-Step qRT-PCR Kits provide:

• SuperScript® III RT for higher temperature cDNA synthesis for greater success with difficult RNA secondary structure
• Platinum® Taq DNA Polymerase with hot-start technology for improved specificity
• LUX™ Fluorogenic Primers (Figure 1) or dual-labeled fluorogenic probes (Figure 2) for superior detection performance
• SYBR® Green I dye for easy and convenient detection (Figure 3)
• Validated performance on multiple real-time instrument platforms, including rotor-based systems

TaqMan™ Gene Expression Cells-to-CT™ Kit Invitrogen™

The TaqMan® Gene Expression Cells-to-CT™ Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification. This kit saves time and offers a simple workflow that is suitable for a few samples or can be easily incorporated into automated, high-throughput applications.

The TaqMan® Gene Expression Cells-to-CT Kit is:

• Complete—optimized workflow includes cell lysis reagents with gDNA removal, RT enzyme mix, buffer, and new TaqMan® Gene Expression Master Mix
• Fast—7-minute sample prep, including DNase treatment, at room temperature
• Easy—lyse samples in a tube or directly in culture plates
• Robust—perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA
• Efficient—contains sufficient reagents to generate 500 real-time PCR results from 100 starting samples

Featuring a unique method for lysing cultured cells while removing genomic DNA and preserving RNA integrity, the TaqMan® Gene Expression Cells-to-CT™ Kit contains reverse transcription (RT) reagents for cDNA synthesis and TaqMan® Gene Expression Master Mix for real-time PCR analysis. TaqMan® primer⁄probe sets are sold separately.

Simple 7-minute sample preparation: part of a complete workflow
Whether you are using plates or tubes, the TaqMan® Gene Expression Cells-to-CT™ Kit, which uses the simple 7-minute sample preparation procedure outlined (see figure), is designed for 10–100,000 cultured cells⁄sample. Cells are washed in PBS and lysed in solution for 5 minutes at room temperature; DNase treatment can be performed simultaneously. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The lysates are now ready for reverse transcription or storage at –20°C. Because samples can be processed directly in culture wells (96 or 384 wells), sample handling and the potential for sample loss or transfer error are minimized, facilitating rapid, high-throughput processing. Unlike old-fashioned, multi-step RNA isolation protocols, no heating, washing, or centrifugation steps are required. The kit greatly simplifies a laborious 30–60 minute process and reduces it to 7 minutes.

The TaqMan® Gene Expression Cells-to-CT™ Kit workflow enables unsurpassed gene expression evaluation with any of the >700,000 TaqMan® Gene Expression Assays. This kit has been extensively tested for specificity with a broad selection of TaqMan® Gene Expression Assays and shows performance equivalent to that obtained with purified RNA (see figure).

Achieve unsurpassed performance and sensitivity
Unlike some competitor kits that limit the amount of lysate in the RT reaction to 5%, the TaqMan® Gene Expression Cells-to-CT™ Kit can accommodate 45% of the total RT reaction volume as cell lysate. Additionally, cDNA can comprise up to 45% of the real-time PCR reaction volume. The large lysate volume in the optimized RT reaction, along with the large cDNA volume in the subsequent real-time PCR using the TaqMan® Gene Expression Master Mix, lead to maximum sensitivity. The master mix amplifies the target precisely and accurately, enabling the detection of small quantities of target, such as transcripts expressed at low levels.

The ability to detect limited target quantities was tested by mixing a constant number of human cells with various numbers of mouse cells. The cell mixtures were prepared using the TaqMan® Gene Expression Cells-to-CT™ and assayed for a mouse-specific and human-specific gene. Comparative data were generated in the same manner with RNA purified by traditional methodology. The data show that RNA from as few as 10 mouse cells was detectable in a background of RNA from 10,000 human cells (see figure). The ability of the TaqMan® Gene Expression Cells-to-CT™ Kit to detect relative low abundance transcripts was equivalent to that of purified RNA; at low levels, it was superior.

Additionally, the performance of the TaqMan® Gene Expression Cells-to-CT™ Kit was compared to competitor lysate kits and to purified RNA. Inputs of 100–100,000 cells⁄lysis reaction were examined. The sensitivity of the TaqMan® Cells-to-CT™ Kit protocol was equivalent to that obtained with purified RNA, and it surpassed competitor lysates (see figure). Furthermore, the lack of inhibition at high cellular inputs and the low variation among technical replicates demonstrate the reliability of this approach for gene expression studies using cultured cells.

Proven performance, proven together
All components of the TaqMan® Gene Expression Cells-to-CT™ Kit have been optimized for consistent and reliable performance. This removes the guesswork involved in assembling separate sample preparation, RT, and real-time PCR kits. And the TaqMan® Gene Expression Cells-to-CT™ Kit has been validated with TaqMan® Gene Expression Assays for added quality assurance.

Accessory product
The TaqMan® Cells-to-CT™ Control Kit was designed for use with the TaqMan® Gene Expression Cells-to-CT™ Kit. This kit contains an endogenous control (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding TaqMan® Gene Expression Assay to monitor the effects of PCR inhibition.

Path-ID™ Multiplex One-Step RT-PCR 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.

Power SYBR™ Green Cells-to-CT™ Kit Invitrogen™

Ambion® Power SYBR® Green Cells-to-CT™ Kit is a fast, easy, and robust method to go directly from cultured cells to real-time PCR results. A breakthrough cell lysis and RNA stabilization technology eliminates the laborious and time-consuming RNA purification process completely. The kit includes sufficient reagents for 400 reactions. Power SYBR® Green Cells-to-CT™ Kit is Powered by AmpliTaq Gold® DNA Polymerase, LD, which provides the highest levels of specificity with standard real-time PCR.

• Includes optimized reagents to work efficiently, right out of the box
• Simple, effective Cells-to-CT™ technology enables sample preparation at room temperature in only 7 minutes, including DNase treatment
• Validated accuracy, exceptional reproducibility, and maximal sensitivity from 10 to 100,000 cells per sample; results equivalent to those from purified RNA

Extraordinary Value
The kit includes reagents for sample preparation integrated into a complete, optimized gene expression workflow including reverse transcription reagents and a high-performance PCR master mix with SYBR® Green.

Extraordinary Ease
The protocol begins with a simple 7-minute sample preparation procedure. First, cells are washed in PBS, and then lysed for 5 minutes at room temperature; DNase treatment can be performed concurrently. The lysis reaction is stopped, and the sample is then ready for reverse transcription or storage at –20°C for up to 5 months. Because samples are processed directly in culture plates (96- or 384-well), sample handling and the potential for sample loss or transfer error are minimized, resulting in higher reproducibility. Following sample preparation, a portion of the cell lysate is added to an RT reaction followed by real-time PCR using the included master mix.

Extraordinary Performance
To demonstrate the performance of the kit, replicate experiments were run using either traditional RNA purification and real-time PCR analysis, or the Power SYBR® Green Cells-to-CT™ Kit. The kit shows equivalent performance to results obtained using purified RNA for analysis of 10 to 100,000 cells. In addition, the sensitivity, efficiency, and dynamic range were shown to be superior to competitor lysate kits. Samples prepared with the kit have been tested for compatibility on various real-time PCR platforms including those from Applied Biosystems®, Roche, and Bio-Rad.

Accessory Product:
The SYBR® Green Cells-to-CT™ Control Kit (SKU# 4402959) contains a Xeno™ RNA control and primer set and is recommended to monitor the efficiency of cell lysis and amplification inhibition in Cells-to-CT™ reactions.

SuperScript™ III Platinum™ One-Step qRT-PCR Kit w/ROX Invitrogen™

The SuperScript III Platinum One-Step qRT-PCR Kit’s proven formula delivers reliable, accurate target detection when it matters most. The convenient one-step format and consistent performance make it ideal for a number of gene expression applications, including infectious disease surveillance. The kit has been validated by several global public health organizations and has long been the choice of the US CDC for infectious disease surveillance.

Features of the SuperScript III Platinum One-Step qRT-PCR Kit include:
• SuperScript III RT for higher temperature cDNA synthesis when addressing difficult RNA secondary structure
• Platinum Taq DNA Polymerase with hot-start technology for improved specificity
• LUX fluorogenic primers and dual-labeled fluorogenic probes for effective detection performance
• ROX reference dye pre-mixed at optimal concentration for Applied Biosystems instruments
• Validated performance on multiple real-time instrument platforms, including rotor-based systems

SuperScript III Platinum One-Step qRT-PCR Kits come in multiple configurations:
SuperScript III Platinum One-Step qRT-PCR Kit with ROX passive reference (this page)
SuperScript III Platinum One-Step qRT-PCR Kit
SuperScript III Platinum SYBR Green One-Step qRT-PCR Kit
SuperScript III Platinum SYBR Green One-Step qRT-PCR Kit with ROX passive reference

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.

Verso 1-step RT-qPCR Kit, with separate ROX vial Thermo Scientific™

The Thermo Scientific Verso 1-step RT-qPCR Kit for probe chemistry offers a quick and simple way to detect mRNA. The Verso RT enzyme delivers high processivity and can be used at temperatures up to 57°C, delivering effective transcription through difficult RNA secondary structures. This version of the kit comes with a separate vial of ROX reference dye.

Features of the Verso 1-step RT-qPCR Kit:

• Eliminate the need for DNase treatments—RT Enhancer (optional) prevents gDNA carry-over
• RT and qPCR steps performed in single tube—convenience and reduced contamination risk
• Use of the entire RT reaction as template for qPCR—improved sensitivity
• Verso RT enzyme is highly sensitive with broad dynamic range—accurate detection of RNA from 1 pg-1 ug
• Verso RT enzyme has a wide working temperature range (42 to 57°C)—improved success with GC-rich and other difficult templates
• Contains an inert blue dye for ease of pipetting during reaction set up

An optional RT Enhancer is also provided and can be used during the RT step to remove any DNA contamination, eliminating the need for DNase I treatment. All Verso 1-step RT-qPCR kits incorporate an inert dye to significantly enhance the contrast between reagent and plastic, making verification of master mix dispensing quick, easy, and foolproof.

In a 1-step approach to RT-qPCR, both the RT step and qPCR steps are performed in a single tube (1-step RT-qPCR). Adopting a 1-step approach is ideal when no cDNA storage is necessary for subsequent experiments. The approach minimizes the risk of contamination and improves reproducibility by reducing sample handling.

Applications

• Gene expression analysis
• RNAi validation
• Microarray validation
• Pathogen detection
• Genetic testing
• Disease research

Related Products
Verso 1-step RT-qPCR Mix, low ROX
Verso 1-step RT-qPCR Mix, ROX
Verso 1-step RT-qPCR Kit, with separate ROX vial

Fast SYBR™ Green Cells-to-CT™ Kit Invitrogen™

Ambion® Fast SYBR® Green Cells-to-CT™ Kit is a fast, easy, and robust method to go directly from cultured cells to real-time PCR results. A breakthrough cell lysis and RNA stabilization technology eliminates the laborious and time-consuming RNA purification process completely. The kit includes sufficient reagents for 400 reactions. The Fast SYBR® Green Cells-to-CT™ Kit contains AmpliTaq® Fast DNA Polymerase, UP, which allows instant hot start, thus minimizing non-specific product formation with fast real-time PCR.

• Includes optimized reagents to work efficiently, right out of the box
• Simple, effective Cells-to-CT™ technology enables sample preparation at room temperature in only 7 minutes, including DNase treatment
• Validated accuracy, exceptional reproducibility, and maximal sensitivity from 10 to 100,000 cells per sample; results equivalent to those from purified RNA

Extraordinary Value
The kit includes reagents for sample preparation integrated into a complete, optimized gene expression workflow including reverse transcription reagents and a high-performance PCR master mix with SYBR® Green.

Extraordinary Ease
The protocol begins with a simple 7-minute sample preparation procedure. First, cells are washed in PBS, and then lysed for 5 minutes at room temperature; DNase treatment can be performed concurrently. The lysis reaction is stopped, and the sample is then ready for reverse transcription or storage at –20°C for up to 5 months. Because samples are processed directly in culture plates (96- or 384-well), sample handling and the potential for sample loss or transfer error are minimized, resulting in higher reproducibility. Following sample preparation, a portion of the cell lysate is added to an RT reaction followed by real-time PCR using the included master mix.

Extraordinary Performance
To demonstrate the performance of the kit, replicate experiments were run using either traditional RNA purification and real-time PCR analysis, or the FastSYBR® Green Cells-to-CT™ Kit. The kit shows equivalent performance to results obtained using purified RNA for analysis of 10 to 100,000 cells. In addition, the sensitivity, efficiency, and dynamic range were shown to be superior to competitor lysate kits. Samples prepared with the kit have been tested for compatibility on various real-time PCR platforms, including those from Applied Biosystems®, Roche, and Bio-Rad.

Accessory Product:
The SYBR® Green Cells-to-CT™ Control Kit (SKU# 4402959) contains a Xeno™ RNA control and primer set and is recommended to monitor the efficiency of cell lysis and amplification inhibition in Cells-to-CT™ reactions.
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