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Synthesize High Yields of Biotinylated aRNA

phosphates during the in vitro transcription (IVT) step, is the preferred label for most commercial microarray platforms. The highly specific and efficient binding of streptavidin-dye conjugates (phycoerythrin,
Cy dyes) to biotinylated RNA/oligonucleotide probe hybrids is the basis of highly sensitive fluorescence detection. Ambion has incorporated biotin labeling reagents into the MessageAmp II Kit to provide a complete microarray aRNA preparation kit, the MessageAmp II-Biotin Enhanced Kit.

Higher Yields of Labeled aRNA; Only 1 Round of Amplification

Both biotin incorporation and further optimization of the IVT reaction has resulted in increased sensitivity of microarray transcript detection. The increased sensitivity allows the use of less input RNA-50 ng of input RNA in a single round of amplification provides enough aRNA for array analysis. Amplified RNA yields derived from nine different tissue and cell sources and four different input amounts are shown in Figure 1. While length of the aRNA synthesized will largely depend on RNA sample quality, the cDNA and IVT reaction conditions in this kit produce longer biotinylated aRNA compared to the market leader. Figure 2 shows comparable Agilent bioanalyzer scan results from labeled aRNA produced with MessageAmp II-Biotin Enhanced Kit and unlabeled aRNA produced with MessageAmp II Kit. The resulting improvements also give the MessageAmp II-Biotin Enhanced Kit a higher number of Present calls than the market leader (Figure 3).


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