Custom RT2 Profiler PCR Arrays

For reliable and sensitive gene expression profiling using laboratory-verified assays

  • Robust and reliable biomarker validation
  • Flexible array format for over 100,000 genes
  • Rigorous QC and QA process for accurate and reliable results
  • Designed for routine use on any real-time cycler

 

 

Custom RT2 Profiler PCR Arrays are sets of optimized real-time PCR primer assays on 96-well plates, 384-well plates, or 100-well discs for a panel of genes related to a specific research area of interest and include appropriate RNA quality controls. Custom RT2 Profiler PCR Arrays enable gene expression analysis with real-time PCR sensitivity and the multi-gene profiling capability of a microarray.

 


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Custom RT2 PCR Array
96-well plate, 384-well plate, or Rotor-Disc 100 including assays, film, or cap strips
330171 Varies
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Custom RT2 PCR Array Additional Plate
Additional Custom RT2 PCR Array plate at time of initial order
330181 Varies
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Custom RT2 Profiler PCR Arrays
96-well plate, 384-well plate, or Rotor-Disc 100 including assays, film, or cap strips
330131 330131
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Create and order

Custom 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.|Simple procedure.|A single gene-specific product in every reaction.|Available plate layouts.|PCR arrays yield highly accurate results.|High reproducibility among different users.|
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 an >80% positive call, even for cytokines expressed at very low levels.||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.||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.|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, is guaranteed to yield single bands of the predicted size without primer-dimers or other secondary products, therefore providing 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 PCR array technology to allow accurate comparison of gene expression across all genes. The unique combination of a 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").

Principle

Custom RT2 Profiler PCR Arrays are reliable tools for studying and verifying gene expression biomarkers in large numbers of biological samples. Whether the biomarkers are from a proprietary collection, literature research, or DNA microarray analysis, QIAGEN’s expertise in real-time PCR detection provides quality, flexibility, and consistency in biomarker validation experiments. QIAGEN’s genomewide coverage provides primers for over 100,000 genes covering the human, mouse, rat, Rhesus macaque, drosophila, and dog genomes.

SYBR® Green based real-time PCR is a powerful technology for studying and validating gene expression biomarkers in large numbers of biological samples. Each 96-well plate, 384-well plate, or 100-well disc PCR array includes SYBR® Green-optimized primer assays. The high-quality primer design and master mix formulation enable the PCR array to amplify gene-specific products simultaneously under uniform cycling conditions.

Rigorous and robust quality assurance (QA) and quality control (QC) procedures covering primer design and quality, specificity, and amplification efficiency provide peace of mind.

This combination provides Custom RT2 Profiler PCR Arrays 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.

Available formats

Custom RT2 Profiler PCR Arrays are available in the following plate layouts (see the tables below and figure "Available plate layouts". Select the genes and the plate format required.

96-well plate or 100-well disc custom qPCR arrays
FormatNumber of plates
(minimum)
8 genes, 12 samples per plate 12
12 genes, 8 samples per plate 12
16 genes, 6 samples per plate 12
24 genes, 4 samples per plate 24
32 genes, 3 samples per plate 24
48 genes, 2 samples per plate 24
96 genes, 1 sample per plate 24
All formats Per additional 12 arrays

384-well plate custom qPCR arrays
FormatNumber of plates
(minimum)
8 genes, 48 samples per plate 6
12 genes, 32 samples per plate 6
16 genes, 24 samples per plate 6
24 genes, 16 samples per plate 6
32 genes, 12 samples per plate 6
48 genes, 8 samples per plate 6
64 genes, 6 samples per plate 6
96 genes, 4 samples per plate 6
128 genes, 3 samples per plate 6
192 genes, 2 samples per plate 24
384 genes, 1 sample per plate 24
All formats Per additional 6 arrays

Arrays are available in additional formats or with special assay designs (position, variants, etc.), please inquire.

Procedure

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

Flexible layout and controls

Custom 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 up to 84 or 370 genes related to a specific research area of interest, plus 5 housekeeping genes. Custom RT2 Profiler PCR Arrays may also include 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

Custom or Modified RT2 PCR Profiler Arrays offer a robust platform for biomarker screening and validation and 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

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Kit Handbooks
2

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For pathway-focused gene expression profiling using real-time RT-PCR
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