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The Ambion® MessageAmp™ II aRNA Amplification Kit is for the amplification of as little as 100 pg of input RNA for GeneChip® analysis. The kit includes sufficient reagents for 20 aRNA amplifications.
• Increased amplification—up to 5,000-fold • Greater sensitivity due to increased yields of full-length cDNA • GeneChip® analysis with as little as 100 ng input RNA • Efficient labeling produces high percent present calls and low 3'/5' ratios • Includes ArrayScript™ RT, a specifically engineered M-MLV reverse transcriptase • Optimized second strand cDNA synthesis reaction
Expression profiling using gene arrays has become the most widely used method for assessing gene expression on a genome-wide/scale. The primary limitation of this technology is the requirement for large amounts of RNA for array hybridization. This hurdle is usually overcome by using T7 RNA Polymerase–driven transcription of cDNA (obtained from total RNA samples) to amplify sufficient amounts of cRNA. This amplification method is the basis of the MessageAmp™ II aRNA Amplification Kit.
An Improved Kit for cRNA Amplification Included in the kit is ArrayScript™ RT, a rationally engineered M-MLV reverse transcriptase, which produces equivalent or higher yields of full-length cDNA compared to other enzymes. The use of ArrayScript™ RT in conjunction with the optimization of the second-strand cDNA synthesis reaction results in increased conversion of mRNA into full-length, double-stranded cDNA templates. In addition, since the in vitro transcription (IVT) uses MEGAscript® technology, the IVT incubation times are shortened. These improvements in the MessageAmp™ II Kit also carry through to two-round reactions, facilitating robust amplification of total RNA from samples as low as 100 pg.
Accessory Products Bio-11-UTP (SKU#AM8450) and Bio-16-UTP (SKU#AM8451)can be used in MessageAmp™ aRNA amplification reactions for target labeling for GeneChip® analysis. The Bio-UTPs are confirmed free of nonspecific endonuclease activity, exonuclease activity, and RNase activity, and are functionally tested using the MessageAmp™ II aRNA Amplification Kit.
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