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Better RNA Amplification

ect incorporation of dye coupled NTPs is inefficient and results in low yields and low specific activity aRNA. This can make the cost of producing aRNA by direct incorporation very high. In addition, incorporation efficiency varies for different labels (e.g. Cy3 NTP vs Cy5 NTP). These problems can be avoided by incorporating an amino allyl NTP into the aRNA and subsequently coupling its reactive amino group with an NHS ester labeled dye (e.g. Cy or other dye). Unlike dye coupled NTPs, amino allyl modified NTPs are incorporated almost as efficiently as unmodified NTPs and are much less expensive than the dye coupled NTPs.

In the Ambion Amino Allyl MessageAmp II protocols, amino allyl UTP is incorporated during the transcription step to produce amino allyl modified aRNA. After a quick purification step, the aRNA is ready for coupling to the NHS ester label of your choice (a variety of inexpensive NHS ester dyes and other nonisotopic labels are available from Amersham Biosciences, Molecular Probes, and Pierce). After dye coupling the reaction is quenched and a final purification is performed, yielding labeled aRNA ready for hybridization.


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Ordering Information Cat# Product Name Size 1751 MessageAmp II aRNA Amplification Kit 20 rxns 1753 Amino Allyl MessageAmp II aRNA Amplification Kit
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