Shop All qRT-PCR Kits

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 100 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.

TaqMan™ RNA-to-CT1-Step Kit (Applied Biosystems™)

The Applied Biosystems TaqMan RNA-to-CT 1-Step Kit delivers consistent RNA target quantification for a wide variety of assays and is validated with Applied Biosystems comprehensive library of TaqMan Gene Expression assays. The kit is recommended for a variety of applications including general gene expression studies, biomarker discovery, and microarray validation.

Features of the TaqMan RNA-to-CT 1-Step Kit include:
• Accurate across a wide dynamic range for dependable performance
• Validated with TaqMan Gene Expression Assays for easy experimental setup
• Consistent performance with a wide variety of targets, including AT-rich, GC-rich and long sequences
• 1-Step format means fewer pipetting steps leading to less chances for errors

Efficiency across a broad range of input concentrations
Maximum flexibility in RNA template input quantity requires optimal real-time PCR efficiency across a broad range of template quantities. Figure 1 shows the excellent amplification efficiency across six orders of magnitude. The result is comparable to that of the TaqMan RNA-to-CT 2-Step Kit, and is a significant improvement in dynamic range and sensitivity over the TaqMan One-Step RT-PCR Master Mix Reagents.

High sensitivity at low target concentration
The sensitivity of TaqMan RNA-to-CT 1-Step Kit was validated using a synthetic RNA template for which copy number is precisely known. Significant sampling error occurs when measuring low quantities of target, so statistical analysis is required for proper evaluation using multiple replicates. Figure 2 shows the expected quantity of target and corresponding mean CT values. Statistical analysis indicates high confidence of sample quantification based on a T-test (Table I), consistent with as few as 10 copies of target*.

*Results are dependent on a variety of factors, including assay design.

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.

TaqPath™ 1-Step RT-qPCR Master Mix, CG (Applied Biosystems™)

TaqPath™ 1-Step RT-qPCR Master Mix, CG, is designed for robust and reproducible one-step pathogen detection and gene expression workflows. The single-tube 4X formulation contains thermostable MMLV reverse transcriptase, dNTPs, UNG, ROX™ dye, and thermostable Fast DNA polymerase, facilitating easy reaction set up—just add user-supplied assay and sample (Figure 1). All manufacturing lots are functionally tested to ensure remarkable lot-to-lot reproducibility for CT consistency and dynamic range across a wide variety of assays. TaqPath™ 1-Step RT-qPCR Master Mix, CG, is a General Purpose Reagent (GPR), manufactured according to applicable 21 CFR requirements and is labeled "For Laboratory Use". With stringent quality coupled with premier performance, TaqPath™ 1-Step RT-qPCR Master Mix, CG, is a superior choice for your diagnostic testing or development needs.

Features of the TaqPath™ 1-Step RT-qPCR Master Mix, CG, include:

• High sensitivity to detect low-copy targets with reproducible CT results
• 6 logs of dynamic range with RNA, DNA, and multiplexing applications*
• Tolerance of inhibitors commonly found in clinical samples
• Labeled "For Laboratory Use"

High Sensitivity
With understanding of the importance of reproducible detection for low-titer pathogen or transcripts in clinical diagnostic testing, TaqPath™ 1-Step RT-qPCR Master Mix, CG, 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 2 shows consistent CT results obtained from three unique lots when detecting 10 copy inputs of RNA target. Figure 3 demonstrates CT lot-to-lot consistency is preserved across multiple assays with different attributes and expression levels.

Wide Dynamic Range Compatible with RNA, DNA, and Multiplexing
TaqPath™ 1-Step RT-qPCR Master Mix, CG, 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 RT-qPCR Master Mix, CG, 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 4 demonstrates the excellent PCR linearity across a 6-log input range for both RNA (Figure 4a) and DNA targets (Figure 4b).

TaqPath™ 1-Step RT-qPCR Master, CG, is compatible with multiplexing reactions, allowing for additional exogenous or endogenous controls or targets to be run simultaneously for quality control or efficiency purposes. Figure 5 demonstrates TaqPath™ 1-Step RT-qPCR Master Mix performance in a triplex reaction. The formulation can be run in either fast or standard cycling conditions with equivalent performance across a wide variety of qPCR platforms.

TaqPath™ 1-Step RT-qPCR Master Mix, CG, has been benchmarked against similar competitor master mixes and demonstrates equivalent or better sensitivity and dynamic range across a variety of targets (Figure 6).

Inhibitor Tolerant
Unlike other master mixes on the market, TaqPath™ 1-Step RT-qPCR Master Mix’s unique proprietary formulation 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 7).

Manufacturing Production and Process Controls
TaqPath™ qPCR Master Mix is manufactured under a strong quality management system that utilizes 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.

General Purpose Reagent
For customers in the USA, TaqPath™ 1-Step RT-qPCR Master Mix is a General Purpose Reagent and "For Laboratory Use".

Cells-to-CT™ 1-Step TaqMan™ Kit (Invitrogen™)

Like other members of the Cells-to-CT™ product family, the Cells-to-CT™ 1-Step TaqMan® Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification.

This new addition to the family includes a TaqMan® 1-Step RT-PCR master mix. The single-tube format of the RT-PCR step streamlines the “sample to answer” workflow more than ever. Combining the simple Cells-to- CT™ lysis technology with the fast cycling capability of the 1-step RT-PCR master mix allows quicker time to results with limited sample handling.

The Cells-to-CT™ 1-Step TaqMan® Kit is:
• Complete—optimized workflow includes cell lysis reagents with gDNA removal and TaqMan® 1-Step RT-PCR Master Mix
• Faster than ever—7-minute sample prep, including DNase treatment, combined with a 28-minute real-time RT-PCR step allows for ~35 minute sample-to-answer workflow
• Simple—10 steps total (sample prep and real-time RT-PCR combined)
• Robust—perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA
• Convenient—contains sufficient reagents to perform 5 real-time RT-PCRs for each cell sample you prepare; this allows you to study multiple targets or perform replicate RT-PCRs

Note: TaqMan® primer/probe sets are sold separately.

Simple 7-minute Sample Preparation, Then Ready for Real-time RT-PCR
Whether you are using plates or tubes, the Cells-to-CT™ 1-Step TaqMan® Kit uses a simple 7-minute sample preparation procedure. The lysis technology is designed for 10–100,000 cultured cells/sample. Cells are washed in PBS and lysed in solution for 5 minutes at room temperature with a simultaneous DNase treatment. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The lysate is now ready to analyze on your real-time PCR instrument. Simply add it to a real-time RT-PCR master mix containing the TaqMan® primer/probe sets of interest. Alternatively, the lysate can be stored 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.

A Complete, Validated Solution
All components of the Cells-to-CT™ 1-Step TaqMan® Kit have been optimized for consistent and reliable performance. This removes the guesswork involved in assembling separate sample preparation and real-time 1-step RT-PCR master mixes. For added quality assurance, the Cells-to-CT™ 1-Step TaqMan® Kit has been validated with TaqMan® Gene Expression assays and shows performance equivalent to that obtained with purified RNA.

Accessory Product
The TaqMan® Cells-to-CT™ Control Kit was designed for use with TaqMan® Cells-to-CT™ products. 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.

EXPRESS One-Step Superscript™ qRT-PCR Kit, with premixed ROX (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 ROX premixed or separate–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, with premixed ROX (this page)
EXPRESS One-Step Superscript qRT-PCR Kit, universal
EXPRESS One-Step SYBR GreenER Kit, with premixed ROX
EXPRESS One-Step SYBR GreenER Kit, universal

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.

TaqMan™ Fast Virus 1-Step Master Mix (Applied Biosystems™)

TaqMan Fast Virus 1-Step Master Mix is designed for fast, highly sensitive real-time RT-PCR even in the presence of challenging PCR inhibitors. The 4X formulation provides enhanced detection of both RNA and DNA viruses and is ideal for multiplex gene expression studies, even with low target input. The TaqMan Fast Virus 1-Step Master Mix provides a single-tube format for uniform handling and processing, while delivering qPCR cycling in <30 minutes, helpful when working in high throughput workflows.

Key product features:
High sensitivity—4X master mix to amplify both RNA and DNA with improved sensitivity
Consistent results—formulated to handle common RT-PCR inhibitors found in blood, stool, and other difficult samples
Simplified application— one-tube, one-step master mix, offering a single run profile with RNA and DNA, which allows for easy mix-and-match of targets on a plate
Target flexibility—works in singleplex, multiplex, and with exogenous or endogenous internal controls
High efficiency—increased qRT-PCR speed on fast and on standard instruments

High sensitivity
Many common research samples with viral nucleic acids have very low levels of target. We have optimized the master mix for high-sensitivity detection of viral targets. The higher-concentration master mix allows you to set up smaller reactions, which means that you can perform fast cycling protocols and obtain at least the same sensitivity as is expected from standard-cycling qPCR. Alternatively, larger sample input amounts can be added to standard reaction volumes for more accurate quantification of low-titer samples (Figure 1).

Consistent results in the presence of inhibitors
Research samples commonly assayed for viruses include blood, dirt, and tissues. Buffer components and proprietary additives in the TaqMan Fast Virus 1-Step Master Mix have been optimized to handle RT-PCR inhibitors (Figure 2) to help ensure consistent performance even with these difficult samples, so that you can be more confident in your results.

Flexibility in targets
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 acid, so you can perform RNA and DNA virus queries next to each other on the same plate using the same handling steps (Figure 3). Additionally, because virus research often includes multiplexed primers and probes and internal reaction controls, we have optimized the master mix to work with multiple target amplicons (Figure 4).

Fast results
The TaqMan Fast Virus 1-Step Master Mix speeds your time to results and maximizes the use of your real-time PCR instruments with a fast protocol. The high-concentration formulation allows for more target nucleic acid sample to be added in the smaller reaction volumes required to run fast protocols. This enables you to maintain sensitivity with low-titer research samples while improving speed and throughput (Figure 5).

TaqMan Fast Virus 1-Step Master Mix provides a reliable and efficient reagent for real-time RT-PCR of virus samples that does not sacrifice accuracy. Its robust performance in the presence of common RT-PCR inhibitors and its convenient and flexible reaction setup allows you to have more confidence in your results.

Cells-to-CT™ 1-Step Power SYBR™ Green Kit (Invitrogen™)

Like other members of the Cells-to-CT™ product family, the Cells-to-CT™ 1-Step Power SYBR® Green Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification.

This new addition to the family includes a Power SYBR® Green 1-Step RT-PCR master mix. The 1-step format of the RT-PCR step streamlines the Cells-to- CT™ "sample to answer" workflow more than ever without compromising sensitivity and specificity and at an extraordinary value.

The Cells-to-CT 1-Step Power SYBR® Green Kit is:
• Complete—optimized workflow includes cell lysis reagents with gDNA removal and Power SYBR® Green 1-Step RT-PCR Master Mix
• Faster—7-minute sample prep, including DNase treatment, versus 30-60 minutes for traditional RNA purification
• Simple—12 steps total (sample prep and real-time RT-PCR combined)
• Robust—perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA
• Affordable—contains sufficient reagents to perform 5 real-time RT-PCRs for each cell sample prepared; study multiple targets per sample at an affordable price

Simple 7-minute Sample Preparation, Then Ready for Real-time RT-PCR
Whether you are using plates or tubes, the Cells-to-CT™ 1-Step Power SYBR® Green Kit uses a simple 7-minute sample preparation procedure. The lysis technology is designed for 10–100,000 cultured cells/sample. Cells are washed in PBS and lysed in solution for 5 minutes at room temperature with a simultaneous DNase treatment. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The lysate is now ready to analyze on your real-time PCR instrument. Simply add it to a real-time RT-PCR master mix containing the primer sets for your genes of interest. Alternatively, the lysate can be stored 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.

Completely Validated, Completely Affordable
All components of the Cells-to-CT™ 1-Step Power SYBR® Green Kit have been optimized for consistent and reliable performance. This removes the guesswork involved in assembling separate sample preparation and real-time 1-step RT-PCR master mixes.

For added quality assurance, the Cells-to-CT™ 1-Step Power SYBR® Green Kit has been validated with a set of primers for common gene targets and shows performance equivalent to that obtained with purified RNA.

When you add it all up, the Cells-to-CT™ 1-Step Power SYBR® Green Kit is the best value for your gene expression experiments. With enough reagents to perform 5 real-time RT-PCRs per sample, the savings are substantial compared to buying master mix and an RNA purification kit separately to do the same work.

Accessory Product
The SYBR® Green Cells-to-CT™ Control Kit was designed for use with SYBR® Green Cells-to-CT™ products. This kit contains an endogenous control primer set (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding primer set to monitor the effects of PCR inhibition.

TaqMan™ PreAmp Cells-to-CT™ Kit (Invitrogen™)

The Ambion® TaqMan PreAmp Cells-to-CT Kit makes it possible to perform gene expression analysis directly from limited or small numbers of cultured cells without RNA purification. The protocol includes an intermediate amplification step (pre-amplification) between reverse transcription and real-time PCR in which cDNA is enriched for up to 100 gene targets of the user’s choice without amplification bias.

• Complete, validated solution—Optimized workflow includes cell lysis reagents, DNase, RT reagents, PreAmp reagents, and TaqMan® Gene Expression Master Mix
• Fast, simple and convenient—Prepare samples in 7 minutes; eliminate tedious RNA isolation
• Extend limited samples—Increase real-time PCR potential by up to 64-fold; study up to 100 genes at once
• Accurate results—Uniform amplification without introducing bias
• Robust performance—Linear detection from 10 to 100,000 cells; results equivalent to purified RNA

Complete, Validated Solution

Featuring a unique method for lysing cultured cells while removing genomic DNA and preserving the RNA integrity, the TaqMan® PreAmp Cells-to-CT™ Kit contains reverse transcription (RT) reagents for cDNA synthesis, TaqMan® PreAmp Master Mix for pre-amplification of up to 100 gene targets, and TaqMan® Gene Expression Master Mix for real-time PCR analysis.

Simple, Convenient 7-Minute Sample Preparation

The TaqMan® PreAmp Cells-to-CT™ Kit incorporates a simple 7-minute sample preparation procedure (Figure 1) for use with 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 concurrently. 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 plates (96 or 384 wells), sample handling and the potential for sample loss or transfer error are minimized, facilitating extension of limited samples. No heating, washing, or centrifugation is required; the kit greatly simplifies a traditionally time-consuming, labor intensive process and reduces it to 7 minutes.

Extension of Limited Samples—Increase Real-Time PCR Potential by up to 64-fold

The integration of the Cells-to-CT™ technology with pre-amplification capability streamlines a multi-step workflow while significantly extending (greater than 60-fold) the number of PCR reactions possible from precious samples, and increases the sensitivity of mRNA detection from small amounts of sample input.

The standard real-time PCR reaction for gene expression analysis starts with the reverse transcription of total RNA to cDNA using random primers and oligo dT, followed by real-time PCR using gene-specific primers and probes (Figure 2). With the TaqMan® PreAmp Cells-to-CT™ Kit, an intermediate amplification step (pre-amplification) between reverse transcription and real-time PCR is performed in which cDNA is enriched for up to 100 gene targets of the user’s choice without amplification bias. The resulting pre-amplified reaction is diluted and serves as the starting material for the subsequent individual TaqMan® Gene Expression Assays using the TaqMan® Gene Expression Master Mix, included in the kit. The TaqMan® PreAmp process effectively extends samples up to 64-fold, enabling analyses previously not possible from limited sample material.

Accurate Results—Equal Amplification without Introducing Bias

The ability to conduct reliable and uniform pre-amplification is critical for gene expression analysis from limited samples. While other methods are available to pre-amplify RNA or cDNA, they use non-specific priming to amplify the starting material, resulting in biased and inaccurate amplification of some transcripts over others. The TaqMan® PreAmp Master Mix, part of the TaqMan® PreAmp Cells-to-CT™ Kit, uses gene specific primers to pre-amplify cDNA resulting in unbiased amplification. The TaqMan® PreAmp Cells-to-CT™ Kit demonstrates unbiased pre-amplification equivalent to purified RNA when compared across 100 different TaqMan Gene Expression Assays. As shown in Figure 3, the pre-amplification reaction does not introduce bias from Cells-to-CT™ lysates nor purified samples; ≥ 92% of the assays were within +⁄- 1.5 ΔΔ CT for pre-amplified vs. un-amplified cDNA from either purified RNA (A) or from Cells-to-CT™ Kit lysates (B).

Robust Performance—Results Equivalent to Purified RNA
Performance of the TaqMan® PreAmp Cells-to-CT™ Kit was evaluated with 100 different TaqMan® Assays utilizing Cells-to-CT™ lysates or purified RNA, either with or without pre-amplification. As shown in Figure 4, the CT values obtained with TaqMan® PreAmp Cells-to-CT™ Kit lysates were equivalent to, and frequently better than CT values obtained using purified RNA, either prior to (A) or after pre-amplification (B). The high level of performance from TaqMan® PreAmp Cells-to-CT™ Kit lysates can be attributed to the preservation of RNA in the starting samples without loss from heating, transfer, or adsorption to matrices.

Linear Detection from 10 to 100,000 Cells

Each step of a workflow for profiling RNA must be robust to work from a few, or many cells. The TaqMan® PreAmp Cells-to-CT™ Kit has been optimized to efficiently generate linearity across 5 logs of cellular input, from 10–100,000 cells per lysis reaction (Figure 5). Furthermore, to maximize sensitivity, the TaqMan® PreAmp Cells-to-CT™ Kit allows up to 45% of the RT reaction volume to be comprised of sample lysate. The TaqMan® PreAmp Cells-to-CT™ Kit provides maximal sensitivity derived from the large lysate volume used in the RT reaction, optimized RT reaction conditions, and sample pre-amplification prior to real-time PCR.

Proven Performance, Proven Together

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

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

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 100 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.

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 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.

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 (11753-100), containing a separate tube of ROX™ Reference Dye, is also available.