The
Full Spectrum Complete
Transcriptome RNA Amplification Kit
makes use of a specially developed Universal
Primer Mixture that uniformly amplifies gene transcripts
using low-cycle PCR. This approach maintains the relative levels of each
transcript in the starting mRNA samples (see data)—even when using starting amounts of RNA as low as 5 ng (see data). The Full Spectrum approach is also faster,
requires fewer steps, and is more convenient than other techniques,
including T7-based in vitro transcription, SMART cDNA amplification, or
Ribo-SPIA amplification.
The Full Spectrum protocol entails just two steps:
Universal Primer Binding and Synthesis of first-strand cDNA.
The Universal Primer preferentially binds to all regions of messenger RNA and then initiates synthesis of the
complementary DNA strand. This step takes approximately 1 hour. Remaining Universal Primer arbitrarily binds to the
first-strand cDNA to prime second-strand synthesis in the following step.
Second-strand cDNA synthesis and amplification.
After an initial 5 min incubation to synthesize the second-strand, cycling commences to amplify the cDNA template.
After 1 hour amplification, you have amplified template—ready for gene-specific PCR.
The amplified product can be used
directly for quantitative PCR using primers specific for any gene
sequence. No purification is required.
