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.
For Research Use Only. Not for use in diagnostic procedures.
Includes Label or Dye
Poly(A+) RNA, Total RNA
aRNA Amplification Kit
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)
Size profile of amplified aRNA.
MessageAmp™ II procedure.
ArrayScript™ Reverse Transcriptase synthesizes intact cDNA products as long as 9 kb.
96-well RNA amplification.
ArrayScript™ Reverse Transcriptase produces up to twice the aRNA yield in MessageAMP™ II: