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Synthesize High Yields of Biotinylated aRNA


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Figure 1. aRNA Yield from Nine Different Sources. Four different amounts of total RNA from nine different sample types were amplified using the MessageAmp II-Biotin Enhanced Kit. Average aRNA yields from triplicate reactions are shown for 4 or 14 hr in vitro transcription (IVT) reactions. These data are useful for determining both the amount of total RNA needed to obtain enough labeled aRNA for array hybridization (typically ~10 g) and the optimal length of IVT incubation. Note that there is a ~3-fold difference in aRNA yield between some samples. With most RNA sources, 50100 ng of input total RNA amplified with the MessageAmp II-Biotin Enhanced Kit using a 14 hr IVT incubation will yield enough labeled aRNA for microarray hybridization.

Figure 2. Electropherogram of Biotin-labeled vs. Unlabeled aRNA. Identical HeLa cell total RNA samples (1000 ng) were amplified with either the MessageAmp II-Biotin Enhanced Kit to produce biotin-labeled aRNA or the MessageAmp II Kit to produce unlabeled aRNA. The IVT reactions were carried out for 4 hr. Equivalent mass amounts from each reaction were analyzed on an Agilent bioanalyzer. The shift in aRNA size from the MessageAmp II-Biotin Enhanced sample results from the incorporation of biotin into the aRNA.

Figure 3. Number of Present Calls Detected on Affymetrix Human Genome Focus Arrays. The indicated amounts of total RNA from HeLa cells was amplified in triplicate using either the Mess
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