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Improving RNA Amplification

alize that all methods will have some inherent bias [8]. Some of this bias can be minimized with a robust protocol and optimized reagents. The MessageAmp II aRNA Amplification Kit provides such a protocol and the novel reverse transcriptase (ArrayScript) for this purpose. We attempted to monitor correlation and standard deviation between two important factors, amplification protocol and RNA input. The data indicate that the MessageAmp II protocol is highly reproducible and can be meaningfully compared to the Affymetrix standard protocol and the original MessageAmp protocol (see sidebar, MessageAmp II Concordance with Original MessageAmp Kit). We hope to have provided some guidelines for amplification expectations and confidence that a shortened IVT incubation time that produces enough cRNA for a microarray will not significantly effect expression data (compared to longer IVT times). The above examples have been an important part of our research, production, and quality control development for procedures that we use to maintain the highest quality reagents for microarray analysis. We plan on continuing such studies for both new product development and improving our line of RNA amplification products.


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