Custom RT² Profiler PCR Arrays

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 for human, mouse, rat, rhesus macaque (compatible with crab-eating macaque (Macaca fascicularis, Cynomolgus monkey), cow, dog, drosophila, horse, pig, rabbit, chicken, zebrafish and CHO cells.

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").

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

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 ΔΔC_T method with normalization of the raw data to either housekeeping genes.

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
• Neuroscience
• Signal transduction pathways
• Cell adhesion and cell migration