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RT² Profiler™ PCR Array System Performance

PCR Arrays stand out in their superb real-time PCR performance. In this section, we present results representative of PCR Array performance based on the following criteria:

  • Sensitivity
  • Specificity
  • Reproducibility
  • Amplification Efficiency

High Sensitivity:
The complete PCR Array System yields a greater-than 85 percent positive call with as little as 25 ng as much as 5 µg of total RNA from a pathway representing genes expressed at a low level (inflammatory cytokines and receptors).

Figure 1: PCR Arrays Let You See More Genes with Less RNA
Different amounts of universal total RNA were characterized using the Human Inflammatory Cytokines and Receptors PCR Array, and the percentage of detectable genes was calculated for each RNA amount.
As little as 25 ng total RNA yields greater than an 80% positive call, even for cytokines expressed at very low levels.

Superb Reproducibility
The complete PCR Array System demonstrates strong correlations across technical replicates, lots, and instruments with average correlation coefficients > 0.99 insuring reliable detection of differences in expression between biological samples.

  050825_1  050825_2  050825_3  050825_4
060111_1 0.993 0.989 0.995 0.992
060111_2 0.994 0.990 0.995 0.992
060111_3 0.992 0.990 0.993 0.992
060111_4 0.993 0.992 0.994 0.992
Figure 2: PCR Arrays Yield Highly Reproducible Results
Four replicate sets of raw threshold data (1-4) obtained by two different scientists (A & B) at two different times (050825 & 060111) on Human Drug Metabolism RT?Profiler PCR Arrays are directly compared. The results demonstrate a high degree of correlation (R2 > 0.990).

Distinct Specificity
The complete 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 thus providing the most accurate real-time PCR results possible.

Figure 3: PCR Arrays Amplify A Single Gene-Specific Product in Every Reaction.
Universal total RNA was characterized on the TGFβ / BMP Signaling Pathway PCR Array, followed by dissociation (melt) curve analysis. PCR Arrays specifically detect individual genes despite the expression of related gene family members in the same RNA sample.

Uniform PCR Amplification Efficiency
One prerequisite for PCR Array technology is the uniform PCR amplification efficiency that allows meaningful comparison of gene expression levels of all genes. The unique combination of SABiosciences' proprietary primer design algorithm and rigorous testing of every primer assay by hand guarantees the high performance of every primer assays on PCR Arrays.

Figure 4: PCR Arrays Yield the Most Accurate Results.
A representative set of assays for 4,000 genes used in RT² PCR Arrays have an average amplification efficiency of 99% with a 95% confidence interval from 90-110%.
Consistently high amplification efficiencies enable PCR Arrays to accurately analyze multiple genes simultaneously utilizing the ΔΔCt method.