Custom RT_ PreAMP Primer Mix

This QIAGEN primer mix is designed for amplification of cDNA templates prior to analysis on Custom RT² Profiler PCR Arrays. Used together with the RT² PreAMP cDNA Synthesis Kit, it enables gene expression analysis using as little as 1 ng total RNA through a proprietary amplification process that increases the amount of cDNA available for downstream PCR array analysis. The amplification process is described as unbiased, yielding highly comparable ΔCT values between preamplified and unamplified cDNA and a high correlation of gene expression fold change results between preamplified and unamplified samples.

Supported 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 cells. Specially designed primer mixes are available for human, mouse, or rat Custom RT² Profiler PCR Arrays. First strand cDNA is synthesized from up to 48 different RNA samples using the RT² PreAMP cDNA Synthesis Kit, then preamplified for a custom gene list. Each first strand cDNA synthesis reaction can be amplified using up to 4 different primer mixes, allowing gene expression analysis on up to 4 different custom PCR arrays. The Side Reaction Reducer included in the synthesis kit eliminates residual primers from preamplification, enabling accurate detection on RT² Profiler PCR Arrays. To complete the PCR array procedure, preamplified templates are mixed with an instrument-specific, ready-to-use RT² SYBR Green Mastermix.

Third-party descriptions characterize the mix as a concentrated primer pool formulated to pre-amplify low-abundance cDNA targets prior to qPCR or array analysis. It enables multiplex pre-amplification to increase sensitivity while preserving relative target abundance and is optimized to minimize amplification bias. The mix is compatible with downstream real-time PCR and profiler arrays and is intended for situations where sample input is limited or when detecting rare transcripts, improving assay success rate for low-copy targets.