The MessageAmp™ II aRNA Amplification Kit contains T7 Oligo(dT) Primer, ArrayScript™ RT, RNase Inhibitor,10X First-Strand Buffer, dNTP Mix, Second-Strand Buffer, DNA Polymerase, RNase H, T7 Enzyme Mix, T7 10X Reaction Buffer, T7 ATP (75 mM), T7 CTP (75 mM), T7 GTP (75 mM), T7 UTP (75 mM), Second Round Primers, and Control RNA (1 mg⁄mL HeLa total RNA) which are all stored at –20°C. The Wash Buffer, cDNA Binding Buffer, aRNA Binding Buffer, aRNA Filter Cartridges, aRNA Collection Tubes, cDNA Filter Cartridges, cDNA Elution Tubes, Nuclease-free Water, and detailed Instruction Manual are stored at room temperature.

描述

The MessageAmp II aRNA Amplification Kit amplifies as little as 100 pg of input RNA for GeneChip analysis. Advantages of the MessageAmp II aRNA Amplification Kit include:

• 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

System overview
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 cDNA 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 (Cat. No. AM8450) and Bio-16-UTP (Cat. No. 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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