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Real-Time PCR and Gene Expression |
Gene expression is a highly regulated process that determines when, where, and under what conditions specific genes are activated within an organism. This regulation is crucial for proper functioning, development, and adaptation. Genes are expressed at different times and locations based on internal factors like cell cycle stages and hormonal signals, as well as external factors such as environmental changes. Transcription factors, enhancers, and silencers play key roles in triggering or preventing gene expression. The products of transcription include mRNA, which serves as a template for protein synthesis, and various non-coding RNAs that regulate gene expression and cellular functions.
TaqMan gene expression assays and arrays are advanced tools used by scientists to measure and analyze gene expression with high specificity and sensitivity. These assays utilize real-time PCR to quantify RNA levels and offer over 2.8 million predesigned primer/probe sets covering more than 30 species. This extensive collection helps allow for precise detection and quantification of gene expression, facilitating research in developmental biology, disease mechanisms, and therapeutic interventions. The data generated from these assays help provide valuable insights into the diverse expression products that determine the growth and health of an organism.
Gene expression applications using real-time PCR
- Pathway analysis
- Biomarker signatures
- eQTL analysis
- Mid- to high-throughput studies using the OpenArray platform
Thermo Fisher Scientific has developed two chemistries for the sensitive, accurate quantification of mRNA using real-time PCR: Applied Biosystems TaqMan probe-based chemistry and Applied Biosystems SYBR green dye-based chemistry.
Learn more:Comparison of TaqMan and SYBR chemistries
Real-time PCR gene expression workflow
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Isolate RNA for real time PCR gene expression
Isolating RNA with methods that preserve RNA integrity and expression profiles is crucial for accurate gene expression analysis. Thermo Fisher Scientific offers a variety of RNA isolation kits tailored to different needs. The Single Cell-to-CT Kit is excellent for single cell analysis, while the Cells-to-CT kits provide a rapid and simple protocol for RT-PCR sample preparation without the need for RNA extraction. For high-quality, intact RNA from any biological material, the TRIzol Reagent is an exceptional choice. Additionally, the mirVana miRNA isolation kits allow for the rapid isolation of small RNAs, helping ensure comprehensive RNA analysis.
Learn more:Automated extraction kits
Design and optimize primers for real time PCR gene expression
Primer Express Software v3.0.1 is a versatile tool for designing primers and probes for gene quantitation and allelic discrimination (SNP) real-time PCR applications, supporting both TaqMan and SYBR Green I dye chemistries. It is compatible with various Thermo Fisher Scientific real-time PCR systems, providing flexibility and ease of use with minimal optimization required. The software offers both automated and manual design options, helping ensure robust assay performance and adherence to assay development guidelines to streamline the design process and help enhance laboratory efficiency.
Reverse transcription for real time PCR gene expression
Convert your RNA into cDNA efficiently across a wide range of RNA concentrations using the SuperScript IV VILO Master Mix. This two-step master mix is optimized for RT-qPCR applications, offering earlier CT values and data reproducibility, even when working with RNA samples that contain inhibitors. For those needing different formats, other SuperScript reverse transcriptase options are also available. Explore these solutions to find a fit for your research needs and help achieve reliable, high-quality results.
Amplify cDNA for real time PCR gene expression
Use TaqMan gene expression assays to amplify cDNA for real-time PCR gene expression analysis for a highly reliable and efficient method. These assays employ a specific pair of unlabeled primers and a TaqMan probe labeled with a fluorescent dye, enabling the precise quantification of target cDNA. By amplifying cDNA, TaqMan assays provide high sensitivity and specificity, allowing researchers to detect even low-abundance transcripts with great accuracy. This method is particularly beneficial for generating consistent and reproducible data across various experimental conditions, making it an indispensable tool in gene expression studies.
TaqMan real-time PCR master mixes are optimized for a wide range of research applications, including gene expression and genotyping. These master mixes provide high sensitivity, specificity, and reproducibility, providing accurate quantification of nucleic acid targets. Compatible with both DNA and RNA targets, TaqMan Real-Time PCR Master Mixes streamline the workflow and enhance the reliability of real-time PCR experiments.
Run qPCR for gene expression using instruments for real-time PCR
QuantStudio Real-Time PCR Systems are advanced platforms designed for high-performance quantitative PCR applications, offering exceptional sensitivity and accuracy. These systems support a variety of throughput needs and applications, from basic research to high-throughput screening, with options such as fixed 96-well and 384-well blocks, as well as other highly flexible systems. With features like intuitive software, remote monitoring, and cloud-enabled analytics, QuantStudio systems help streamline workflows and enhance data management for researchers.
Analyze data for real time PCR gene expression
Thermo Fisher Scientific offers a suite of software tools for analyzing real-time PCR gene expression data with high precision and ease. These tools provide robust features for CT analysis, relative quantitation, and advanced data visualization, helping ensure accurate interpretation and reliable results for gene expression.
Explore:ExpressionSuite software
Isolate RNA for real time PCR gene expression
Isolating RNA with methods that preserve RNA integrity and expression profiles is crucial for accurate gene expression analysis. Thermo Fisher Scientific offers a variety of RNA isolation kits tailored to different needs. The Single Cell-to-CT Kit is excellent for single cell analysis, while the Cells-to-CT kits provide a rapid and simple protocol for RT-PCR sample preparation without the need for RNA extraction. For high-quality, intact RNA from any biological material, the TRIzol Reagent is an exceptional choice. Additionally, the mirVana miRNA isolation kits allow for the rapid isolation of small RNAs, helping ensure comprehensive RNA analysis.
Learn more:Automated extraction kits
Design and optimize primers for real time PCR gene expression
Primer Express Software v3.0.1 is a versatile tool for designing primers and probes for gene quantitation and allelic discrimination (SNP) real-time PCR applications, supporting both TaqMan and SYBR Green I dye chemistries. It is compatible with various Thermo Fisher Scientific real-time PCR systems, providing flexibility and ease of use with minimal optimization required. The software offers both automated and manual design options, helping ensure robust assay performance and adherence to assay development guidelines to streamline the design process and help enhance laboratory efficiency.
Reverse transcription for real time PCR gene expression
Convert your RNA into cDNA efficiently across a wide range of RNA concentrations using the SuperScript IV VILO Master Mix. This two-step master mix is optimized for RT-qPCR applications, offering earlier CT values and data reproducibility, even when working with RNA samples that contain inhibitors. For those needing different formats, other SuperScript reverse transcriptase options are also available. Explore these solutions to find a fit for your research needs and help achieve reliable, high-quality results.
Amplify cDNA for real time PCR gene expression
Use TaqMan gene expression assays to amplify cDNA for real-time PCR gene expression analysis for a highly reliable and efficient method. These assays employ a specific pair of unlabeled primers and a TaqMan probe labeled with a fluorescent dye, enabling the precise quantification of target cDNA. By amplifying cDNA, TaqMan assays provide high sensitivity and specificity, allowing researchers to detect even low-abundance transcripts with great accuracy. This method is particularly beneficial for generating consistent and reproducible data across various experimental conditions, making it an indispensable tool in gene expression studies.
TaqMan real-time PCR master mixes are optimized for a wide range of research applications, including gene expression and genotyping. These master mixes provide high sensitivity, specificity, and reproducibility, providing accurate quantification of nucleic acid targets. Compatible with both DNA and RNA targets, TaqMan Real-Time PCR Master Mixes streamline the workflow and enhance the reliability of real-time PCR experiments.
Run qPCR for gene expression using instruments for real-time PCR
QuantStudio Real-Time PCR Systems are advanced platforms designed for high-performance quantitative PCR applications, offering exceptional sensitivity and accuracy. These systems support a variety of throughput needs and applications, from basic research to high-throughput screening, with options such as fixed 96-well and 384-well blocks, as well as other highly flexible systems. With features like intuitive software, remote monitoring, and cloud-enabled analytics, QuantStudio systems help streamline workflows and enhance data management for researchers.
Analyze data for real time PCR gene expression
Thermo Fisher Scientific offers a suite of software tools for analyzing real-time PCR gene expression data with high precision and ease. These tools provide robust features for CT analysis, relative quantitation, and advanced data visualization, helping ensure accurate interpretation and reliable results for gene expression.
Explore:ExpressionSuite software
Tools for performing gene expression using real-time PCR (qPCR) or digital PCR
TaqMan gene expression assays provide outstanding versatility to suit various experimental needs allowing scientists to streamline their gene expression studies and obtain high-quality data with ease.
Amplify 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 the sample. This method can be used to help enable single-cell studies.
Explore:TaqMan PreAmp Master Mix
Thermo Fisher Scientific's one-step RT-qPCR reagents streamline the reverse transcription and PCR amplification processes into a single reaction, helping enhance convenience and helping reduce the risk of contamination. Reagents, such as the TaqMan Fast Virus 1-Step and TaqPath DuraPlex 1-Step RT-qPCR master mix, offer high sensitivity and reproducibility, making them excellent for detecting and analyzing low-abundance RNA targets in various research applications.
Learn more: TaqMan Fast Virus 1-Step master mix for qPCR
Thermo Fisher Scientific's digital PCR technology, exemplified by the Applied Biosystems QuantStudio Absolute Q Digital PCR System, offers precise and sensitive absolute quantification of nucleic acids, making it excellent for detecting rare mutations and target sequences in gene expression studies.
Learn more:Digital PCR
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