Custom RT 2 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 RT 2 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 RT 2 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 RT 2 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 RT 2 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. Rotor-Gene and 96-well plate or 100-well disc custom qPCR arrays Format Number 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 array 384-plate custom qPCR arrays Format Number of plates (minimum) 8 genes, 48 samples per plate 6 12 genes, 32 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 array 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 RT 2 Profiler PCR Arrays are compatible with all QIAGEN, ABI, Bio-Rad, Eppendorf, Roche and Stratagene instruments. Flexible layout and controls Custom RT 2 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 RT 2 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.
