When, where, and under what conditions are genes expressed? What triggers, or prevents, this expression? Scientists are discovering the surprisingly wide range of products of transcription and translation, and how these different expression products determine the growth and health of an organism.

Common gene expression research applications include:

Real-Time PCR
  • Quantitation of gene products
  • Microarray validation
  • Pathway analysis
  • Developmental biology
  • Quality control and assay validation
  • siRNA/RNAi experiments
  • Low-fold copy number discrimination
  • Mid-to high-throughput profiling using the OpenArray® platform
Digital PCR
  • Absolute quantification of nucleic acid standards & next-generation sequencing libraries
  • Rare target detection
  • Enrichment and separation of mixtures

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We have developed three methods for sensitive, accurate quantification of mRNA using real-time PCR: TaqMan® probe-based analysis, SYBR® Green dye-based analysis, and digital PCR.


Need help choosing the right gene expression solution? Try our configuration tool

Comparison of Real-Time PCR Gene Expression Analysis Techniques


Workflow Steps TaqMan® Gene Expression SYBR® Green Dye Gene Expression
Step 1: Isolate RNA

Isolate RNA using methods that preserve RNA integrity and expression profiles

Isolate RNA using methods that preserve RNA integrity and expression profiles

Step 2: Design & Optimize Primers

Not needed, TaqMan® assays have passed our industry leading bioinformatics design criteria

Critical to optimize your primer design to reduce the risk of amplifying non-target sequences and ensure an efficient PCR experiment

Step 3: Reverse Transcription

Convert your RNA into cDNA, over a wide range of RNA concentrations

Convert your RNA into cDNA, over a wide range of RNA concentrations

Step 4: Amplify cDNA

Amplify your cDNA using TaqMan® Assays & TaqMan® master mix. Once you add your cDNA, these products contain all of the enzymes, buffers, primers and probe necessary to perform TaqMan® real-time PCR analysis

Amplify your cDNA using SYBR® Green master mix & custom primers. Together with your cDNA, these products contain all of the enzymes, buffers, primers and probe necessary to perform SYBR Green Real-Time PCR analysis.

Step 5: Run Reactions
Step 6: Analyze Data

Alternatives & Options When Performing Real-Time PCR Gene Expression

We have several options to enhance performance or simplify the steps in performing gene expression studies with real-time PCR.

  • Pre-amplification for limited samples. TaqMan® PreAmp Master Mix can be used to amplify your cDNA before using it for the real-time PCR reaction. It will increase the amount of cDNA available for subsequent reactions without creating bias in the relative amount of cDNA represented in your sample. This method can be used to help enable single-cell studies.
  • One-step RT-PCR reagents: By combining reverse transcription (RT) and real-time PCR amplification (qPCR) reagents into a single reaction, you are able to reduce the number of hands-on manipulations required as well as the total time to process your samples and get data. It is ideal for situations where many samples are being screened for a few targets, or when automating your experiments with liquid handling systems. One potential drawback is that all of the cDNA is used in the qPCR, so none is left over for later use.
  • Cells-to-CT™ Kits and other integrated sample preparation/real-time PCR products eliminate the need for nucleic acid purification, creating much simpler experimental protocols and reducing the effort for sample preparation to just a few minutes.
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Regulatory Status: For Research Use Only. Not for use in diagnostic procedures.