PCR-Based Drug Discovery Assays - Real-Time PCR Kits

TaqMan™ Gene Expression Master Mix (Applied Biosystems™)

Get precise and reliable real-time qPCR quantification over nine logs of linear dynamic range with TaqMan Gene Expression Master Mix. TaqMan Gene Expression Master Mix is an optimized 2X mix that contains all of the components, excluding the template and primers, for sensitive detection down to one copy of target. Extended benchtop stability provides consistent results for automated liquid-handling systems, making it the choice for high-throughput sample processing, overnight or over the weekend. TaqMan Gene Expression Master Mix has been validated with TaqMan gene expression assays and Applied Biosystems real-time systems, providing precise quantification for a variety of gene expression–based real-time qPCR applications, including:
• Rare transcript detection
• Gene expression analysis
• Pathogen detection and viral load quantification
• Co-amplification of two targets
• Specific detection of gene family members

TaqMan Gene Expression Master Mix contains AmpliTaq Gold DNA Polymerase, UP (Ultra Pure), for hot start activation and improved detection of bacterial targets, a blend of dNTPs with dTTP/dUTP and uracil-DNA glycosylase (UDG) to minimize carry-over PCR contamination, and a passive internal reference based on proprietary ROX dye for superb precision on Applied Biosystems real-time PCR instruments.

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 Gene Expression Master Mix and added additional capabilities for your gene expression analysis.

TaqMan™ Universal PCR Master Mix (Applied Biosystems™)

TaqMan Universal PCR Master Mix 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
• AmpErase UNG protects against subsequent re-amplification from PCR products containing dU to minimize carry-over contamination
• 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.

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.

Protein Thermal Shift™ Starter Kit

Applied Biosystems® Protein Thermal Shift™ kits allow users to employ a dye that binds to exposed hydrophobic residues to monitor the thermal stability of proteins using a real-time PCR instrument in order to identify buffer conditions that stabilize the protein of interest or screen libraries of ligands for compounds that bind the protein of interest.

Save Time and Money with Your Sample Screening
This solution significantly reduces the time and cost associated with screening samples relative to currently available methods. Protein Thermal Shift™ products that are run on our real-time PCR systems enable researchers to inexpensively and efficiently:

• Screen samples for relative protein thermal stability
• Screen molecules (e.g. small molecule drug candidates, fragment-based libraries, and antibodies) for ligand binding to proteins in a high-throughput manner
• Screen samples for stability changes after protein point mutations or modifications
• Systematically identify suitable⁄optimal buffer conditions for the measurement of protein-ligand interactions, production of stable protein preparations, protein crystallization, and storage of proteins
• Quickly QC protein preps to ensure protein integrity and⁄or purity

Two Kits to Choose From
The Protein Thermal Shift™ Dye Kit contains the dye and buffer for performing protein thermal shift experiments, enough for up to 2000 reactions. The Protein Thermal Shift™ Starter Kit contains the Dye Kit plus a control protein and control ligand for test experiments and troubleshooting. The Protein Thermal Shift™ Starter Kit components are packaged in two boxes, one to be stored at -20°C and the other at 4°C, for maximum flexibility in reagent formulation.

Simplify Analysis with Protein Thermal Shift™ Software
Our complete Protein Thermal Shift™ solution includes analysis software which is compatible with run files generated from five of our real-time PCR systems: StepOne™ Real-Time PCR System, StepOnePlus™ Real-Time PCR system, 7500 Fast Real-Time PCR System, and ViiA™7 Real-Time PCR System.

Protein Thermal Shift™ Dye Kit

Applied Biosystems® Protein Thermal Shift™ kits allow users to employ a dye that binds to exposed hydrophobic residues to monitor the thermal stability of proteins using a real-time PCR instrument in order to identify buffer conditions that stabilize the protein of interest or screen libraries of ligands for compounds that bind the protein of interest.

Save Time and Money with Your Sample Screening
This solution significantly reduces the time and cost associated with screening samples relative to currently available methods. Protein Thermal Shift™ products that are run on our real-time PCR systems enable researchers to inexpensively and efficiently:

• Screen samples for relative protein thermal stability
• Screen molecules (e.g. small molecule drug candidates, fragment-based libraries, and antibodies) for ligand binding to proteins in a high-throughput manner
• Screen samples for stability changes after protein point mutations or modifications
• Systematically identify suitable⁄optimal buffer conditions for the measurement of protein-ligand interactions, production of stable protein preparations, protein crystallization, and storage of proteins
• Quickly QC protein preps to ensure protein integrity and⁄or purity

Two Kits to Choose From
The Protein Thermal Shift™ Dye Kit contains the dye and buffer for performing protein thermal shift experiments, enough for up to 2000 reactions. The Protein Thermal Shift™ Starter Kit contains the Dye Kit plus a control protein and control ligand for test experiments and troubleshooting. The Protein Thermal Shift™ Starter Kit components are packaged in two boxes, one to be stored at -20°C and the other at 4°C, for maximum flexibility in reagent formulation.

Simplify Analysis with Protein Thermal Shift™ Software
Our complete Protein Thermal Shift™ solution includes analysis software which is compatible with run files generated from five of our real-time PCR systems: StepOne™ Real-Time PCR System, StepOnePlus™ Real-Time PCR system, 7500 Fast Real-Time PCR System, and ViiA™7 Real-Time PCR System.

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

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.

TaqMan™ PreAmp Master Mix Kit (Applied Biosystems™)

Stretch your precious samples into more real-time PCR experiments with TaqMan® PreAmp Master Mix Kit. Applied Biosystems® TaqMan® PreAmp Master Mix Kit contains TaqMan® PreAmp Master Mix and TaqMan® Gene Expression Master Mix. The two work hand in hand to provide a seamless workflow for preamplification and quantitation of mRNA.

The TaqMan® PreAmp Master Mix Kit lets you:

• Amplify cDNA targets equally without introducing bias
• Analyze mRNA from any precious sample such as laser capture microdissections, needle biopsies, and formalin-fixed paraffin-embedded tissues (FFPE)
• 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 Pre-amplification Bias
In the past, preamplification kits and techniques often resulted in uneven amplification of targets and inaccurate or biased data (as shown by low correlation coefficients between amplified and unamplified samples). TaqMan® PreAmp Master Mix amplifies your small starting samples with no bias and provides extremely high correlation between amplified and unamplified cDNA.

Optimized Workflow and Results
TaqMan® PreAmp Master Mix and TaqMan® Gene Expression Master Mix work in conjunction to provide an optimal workflow for analysis of small starting samples. TaqMan® Gene Expression Master Mix provides reliable quantification over 9 logs of linear dynamic range and has been validated with TaqMan® Assays.