RT_ PreAMP Pathway Primer Mix

QIAGEN offers this RT² PreAMP Pathway Primer Mix as part of its Assay Technologies consumables portfolio, designed for RNA expression applications. The primer mix enables gene expression analysis using as little as 1 ng total RNA through a proprietary amplification process that increases the amount of cDNA for subsequent PCR array analysis. It is used for amplification of cDNA templates prior to analysis on RT² Profiler PCR Arrays, and primer mixes are available for all cataloged human, mouse, and rat RT² Profiler PCR Arrays.

Suitable starting sample types include fine needle biopsy and laser captured microdissection samples, stem cell clusters or embryoid bodies, and fluorescence-activated cell sorter (FACS) generated cell populations. First strand cDNA is first synthesized from up to 12 different RNA samples using the RT² PreAMP cDNA Synthesis Kit. The cDNA is then preamplified for a pathway-specific set of genes, and each first strand cDNA synthesis reaction can be amplified using up to 4 different RT² PreAMP Pathway Primer Mixes, allowing gene expression analysis on up to 4 different RT² Profiler PCR Arrays. The Side Reaction Reducer included in the RT² PreAMP cDNA Synthesis Kit eliminates residual primers from preamplification, enabling accurate detection on RT² Profiler PCR Arrays. More genes are detected after preamplification using the mix, and the amplification process is unbiased, yielding highly comparable ΔCT values between preamplified and unamplified cDNA and a high correlation of gene expression fold change results.

To complete the PCR array procedure, preamplified templates are mixed with an instrument-specific, ready-to-use RT² SYBR Green Mastermix. Care should be taken to minimise DNA contamination, which can artificially inflate the SYBR Green signal and produce skewed gene expression profiles or false-positive signals; the most common source is PCR products from previous experiments.