Access the Entire mRNA Transcriptome with Full Spectrum™ RNA Amplification


* Maintain complete mRNA sequence even from degraded RNA
* Excellent results using degraded RNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples Application Note
* Read the Genetic Engineering News Full Spectrum Highlight GENews Article
* Uniform amplification across the whole RNA transcript with no bias toward the 3’- or 5’-ends
* High fidelity maintenance of relative levels of each mRNA species
* Uniform transcript amplification enables you to study alternative mRNA splicing
* Amplify poly A- RNA, e.g. Histone mRNAs such as H1a
* Amplify viral RNA, many of which lack poly A tails
* True one-tube approach that produces amplified template in just 3 hours

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The Full Spectrum™ Complete Transcriptome RNA Amplification Kit provides a superior approach to amplify RNA for expression analysis. In addition to robustly and reliably amplifying difficult RNA from valuable samples in a way that maintains the relative levels of each transcript, the Full Spectrum approach ensures that all regions of the transcripts are present in the amplified products. Unbiased amplification also means that Full Spectrum samples can be used for analysis of gene families and gene transcripts that display splicing variations. Splicing variations may occur in any region of the message, and it is estimated that approximately 40% of genes have multiple splicing variants (Brett, et al, FEBS, 474: 83-86, 2000). This feature also makes Full Spectrum amplification particularly suitable for the preparation of samples for microarray analysis. Starting with just nanogram quantities of RNA, the system provides enough amplified template for quantitative PCR expression analysis of 100-200 different transcripts (depending on your PCR reaction volume).

The degradation of RNA has, until now, made it difficult to reliably amplify enough usable material for expression analysis. Most amplification procedures, such as T7 in vitro transcription and Ribo-SPIA™ RNA amplification, rely on poly-A priming and produce product that is highly biased toward the 3’-end of transcripts. These amplification methods do not typically amplify anything beyond the last 600 to 1,000 bases of the transcript. Other amplification methods leave much of the mRNA inaccessible to analysis, while. Competitor’s 3’-biased approaches are particularly problematic when amplifying degraded RNA derived from formalin-fixed, paraffin-embedded (FFPE) tissues, where they typically cannot produce usable product. Full Spectrum technology allows robust gene expression analysis.

The Full Spectrum™ RNA Amplification Kit makes use of a specially developed Universal Primer that robustly and uniformly amplifies all regions of gene transcripts using low-cycle PCR. The Universal Primer is composed of a proprietary, non-degenerate primer that binds to, and primes synthesis from numerous specific sites found throughout mRNA sequences. The combination of this primer mix and low-cycle PCR (typically < 20 cycles) produces uniform amplification of gene transcripts so the relative levels of each transcript in the starting mRNA sample are maintained—even when using starting amounts of RNA as low as 20 ng. This unique approach is particularly robust for amplifying degraded RNA, making the Full Spectrum method the obvious choice for amplifying RNA from all sources, including FFPE tissues.

Comparison of the most commonly-used RNA amplification methods