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MessageAmp II: Use Less RNA for Array Analysis

und of amplification with MessageAmp II generates enough aRNA for array analysis using Affymetrix GeneChip Arrays. Other commercially available products for RNA amplification require two rounds of amplification at this level of RNA input.

Together, these improvements result in more efficient conversion of mRNA into longer double-stranded cDNA templates. This, in turn, enables shorter in vitro transcription (IVT) reaction times and reliable expression analysis from small RNA samples (less than 1 g of total RNA). The MessageAmp II improvements also have an impact when two rounds of amplification are employed, allowing more robust amplification of total RNA amounts from 10 ng to as little as 100 pg.

Here we provide aRNA yield data and guidelines for the number of amplification rounds needed based on the amount of input total RNA and the RNA source used in the reaction.


Factors Affecting cRNA Yield
Factors such as tissue source and disease state, mRNA content (0.13%), and starting amount and quality of total RNA will all affect the yield of aRNA from any amplification-labeling procedure. Figure 1 shows typical yields of aRNA amplified from total RNA isolated from six different tissues using the MessageAmp II aRNA Amplification Kit. RNA input amounts ranging from 503000 ng were tested from each source. These data show some general trends found to be reproducible. For example, while aRNA yield generally increases with increasing amounts of input RNA, addi
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