MessageAmp™ II aRNA Amplification Kit

Catalog number: AM1751

Ambion™  Related applications: Microarray Analysis

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Description

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:

• 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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 20 reactions
Includes Label or Dye: No
Reverse Transcriptase: ArrayScript™ Reverse Transcriptase
Sample Type (Specific): Total RNA, Poly(A+) RNA

Contents & storage

• 60 µl T7 Oligo(dT) Primer (-20°C)
• 22 µl ArrayScript™ Reverse Transcriptase (-20°C)
• 22 µl RNase Inhibitor (-20°C)
• 42 µl 10X First Strand Buffer (-20°C)
• 170 µl dNTP Mix (-20°C)
• 210 µl 10X Second Strand Buffer (-20°C)
• 42 µl DNA Polymerase (-20°C)
• 22 µl RNase H (-20°C)
• 84 µl T7 Enzyme Mix (-20°C)
• 84 µl T7 10X Reaction Buffer (-20°C)
• 84 µl each T7 ATP, CTP, GTP, and UTP (-20°C)
• 40 µl Second Round Primers (-20°C)
• 10 µl Control RNA 1 mg/ml (-20°C)
• 1.75 ml Nuclease-free Water (any temperature)
• 30 ml Wash Buffer (4°C or room temperature)
• 7 ml cDNA Binding Buffer (room temperature)
• 9 ml aRNA Binding Buffer (room temperature)
• 20 aRNA Filter Cartridges (room temperature)
• 40 aRNA Collection Tubes (room temperature)
• 20 cDNA Filter Cartridges + Tubes (room temperature)
• 20 cDNA Elution Tubes (room temperature)
• 10 ml Nuclease-free Water (any temperature)

Documents

Manuals & protocols

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