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RT2 Profiler PCR Arrays

For pathway-focused gene expression analysis using laboratory-verified assays

  • Profile 84 or 370 related genes simultaneously
  • Expert-designed panels target the most relevant genes
  • Laboratory-verified assays eliminate tedious optimization
  • Simple procedure enables routine use with any real-time PCR instrument
  • Complimentary online tools make data analysis quick and easy
RT² Profiler PCR Arrays enable quick, reliable gene expression analysis of 170+ pathways, allowing researchers to spend less time at the bench and more time interpreting results. The arrays are pathway-focused panels of laboratory-verified qPCR assays, with integrated, patented controls to ensure a successful experiment every time. PhD-trained application specialists are available to provide technical support for the arrays, and each array can also be modified to suit unique experimental needs. For biomarker discoveries, we offer Pathway Plus PCR arrays and all arrays listed below for the Fluidigm BioMark Real-Time PCR System. 
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RT2 Profiler PCR Arrays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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Positive results with as little as 25 ng RNA.
Different amounts of universal total RNA were characterized using the Human Inflammatory Cytokines and Receptors RT2 Profiler PCR Array and the percentage of detectable genes calculated for each amount. As little as 25 ng total RNA yields a >80% positive call, even for cytokines expressed at very low levels.
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Simple procedure.
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A single gene-specific product in every reaction.

Universal total RNA was characterized using the TGFβ/BMP Signaling Pathway RT2 Profiler PCR Array, followed by dissociation (melting) curve analysis. The PCR array specifically detects individual genes, despite the expression of related gene-family members in the same RNA sample.

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PCR arrays yield highly accurate results.
A representative set of assays for 4000 genes used in RT2 Profiler PCR Arrays have an average amplification efficiency of 99% with a 95% CI from 90–110%. Consistently high amplification efficiencies enable PCR arrays to accurately analyze multiple genes simultaneously utilizing the ΔΔCT method.
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High reproducibility among different users.
4 replicate sets of raw threshold data (1–4) obtained by 2 different scientists (A and B) at 2 different times using Human Drug Metabolism RT2 Profiler PCR Arrays were directly compared. The results demonstrate a high degree of correlation (R2 >0.990).
Performance
Sensitivity

With the sensitivity of the RT2 First Strand Kit, as little as 1 ng or as much as 5 µg of total RNA per array plate provides greater than 80% present call rates (see figure "Positive results with as little as 25 ng RNA").

Reproducibility

The complete PCR array system demonstrates strong correlations across technical replicates, lots, and instruments with average correlation coefficients >0.99, ensuring reliable detection of differences in expression between biological samples (see figure "High reproducibility among different users").

Specificity

The PCR array system, with high-quality input RNA, yields single bands of the predicted size without primer-dimers or other secondary products, therefore providing the highly accurate real-time PCR results (see figure "A single gene-specific product in every reaction").

Uniform PCR amplification efficiency

Uniform PCR amplification efficiency is required for the PCR array technology to allow accurate comparisons of gene expression across all genes and all samples. The unique combination of our proprietary primer design algorithm and rigorous testing of every primer assay guarantees the high performance of every primer assay on PCR arrays (see figure "PCR arrays yield highly accurate results").

A guaranteed system

RT² Profiler PCR Arrays are tested and optimized in combination with the RT² SYBR Green qPCR Mastermixes and the RT² First Strand Kit. This testing means that RT² Profiler PCR Array performance is guaranteed when all three of these components are used together.

Principle

RT2 Profiler PCR Arrays are reliable tools for analyzing the expression of a focused panel of genes. Each 96-well plate, 384-well plate or 100-well disc PCR array includes SYBR® Green-optimized primer assays for a thoroughly researched panel of relevant, pathway- or disease-focused genes. RT2 Profiler PCR Arrays can also be customized to contain a panel of genes tailored to your specific research interests. The high-quality primer design and RT2 SYBR® Green qPCR Mastermix formulation enable the PCR array to amplify 96 or 384 different gene-specific products simultaneously under uniform cycling conditions.

This combination provides the RT2 Profiler PCR Array with the specificity and the high amplification efficiencies required for accurate real-time SYBR® Green results. PCR arrays are easy to use in any research laboratory.

RT2 Profiler PCR Arrays are sensitive enough for use with RNA prepared from regular samples (0.1–5 µg RNA), FFPE samples, and small samples (1–100 ng RNA).

Procedure

Simply mix the cDNA template with the appropriate ready-to-use PCR mastermix, aliquot equal volumes to each well of the same plate, and then run the real-time PCR cycling program (see flowchart "Simple procedure"). RT2 Profiler PCR Arrays are compatible with all QIAGEN, ABI, Bio-Rad, Eppendorf, Roche, and Stratagene instruments.

Flexible layout and controls

RT2 Profiler PCR Arrays are available in 96-well plate, 384-well plate, and 100-well disc formats, and are used to monitor the expression of 84 or 370 genes related to a disease state or pathway, plus 5 housekeeping genes. Each RT2 Profiler PCR Array also includes control elements for:

Data normalization  
Genomic DNA contamination detection
RNA sample quality 
General PCR performance

Easy-to-use data analysis

Data can be analyzed using an easy-to-use Excel-based data analysis template or Web-based software. Data analysis is based on the ΔΔCT method with normalization of the raw data to either housekeeping genes.

Applications

RT² PCR Profiler Arrays can be used in all areas of biological and medical research, including:

  • Cancer Inflammation and cytokine profiling
  • Stem cells
  • Neuroscience
  • Signal transduction pathways
  • Cell adhesion and cell migration
  • Biomarker screening and Validation

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Kit Handbooks (4)

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For cDNA synthesis from 96 samples
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パスウェイ特異的遺伝子の発現をリアルタイムRT-PCR を用いてプロファイリング
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For pathway-focused gene expression profiling using real-time RT-PCR
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Instrument Technical Documents (2)
For pathway-focused gene expression analysis
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For gene expression and genomic analysis
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References
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