RT² PreAMP cDNA Synthesis Kit

The RT² PreAMP cDNA Synthesis Kit is designed for preamplification of cDNA from nanogram amounts of RNA (1–100 ng) specifically for analysis using RT² Profiler PCR Arrays.

RT² PreAMP technology utilizes multiplex, tandem PCR to preamplify gene-specific cDNA with minimal bias. The RT² PreAMP cDNA Synthesis Kit is designed for preamplification of first strand cDNA from 1–100 ng total RNA.
Following standard reverse transcription, the kit works with an array-specific RT² PreAMP Pathway Primer Mix to preamplify an array-specific set of targets from the cDNA before gene expression analysis using an RT² Profiler PCR Array. Each RNA sample will generate sufficient cDNA for gene expression analysis on up to 4 different RT² Profiler PCR Arrays.

The procedure comprises 2 simple steps:

• First strand cDNA synthesis: The kit can be used to synthesize cDNA from up to 12 different RNA samples. The built-in external RNA control template, which is detected by the reverse transcription control (RTC) assays in the RT² Profiler PCR Arrays, allows the detection of reverse transcription inhibitors and ensures efficient first strand synthesis.
• Preamplification of cDNA for pathway-specific genes: Each first strand cDNA synthesis reaction from 1–100 ng of total RNA can be amplified using up to 4 different sets of PCR array-specific primer mixes, allowing gene expression analysis on up to 4 different RT² Profiler PCR Arrays. The Side Reaction Reducer included in the kit eliminates the residual primers from preamplification, enabling accurate detection on PCR arrays.

To complete the PCR array procedure, the preamplified templates are mixed with an instrument-specific, ready-to-use RT² SYBR^® Green qPCR Mastermix.

cDNA templates generated using the RT² PreAMP cDNA Synthesis Kit in conjunction with RT² PreAMP Pathway Primer Mixes are ready for gene expression profiling using RT² Profiler PCR Arrays.