Invitrogen™ MessageAmp™ II aRNA Amplification Kit

Amplifies as little as 100pg of input RNA for GeneChip™ analysis

Marke:  Invitrogen™ AM1751

Artikelnummer. 10056234

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

Beschreibung

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 100pg.

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.

• Increased amplification: up to 5,000-fold
• Greater sensitivity due to increased yields of full-length cDNA
• GeneChip analysis with as little as 100ng 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 Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, RNA Amplification

Bestellinformation

Shipping Condition: Approved for shipment on Wet or Dry Ice

Spezifikation

• Product Type: aRNA Amplification Kit
• Includes Label or Dye: No
• Sample Type: Poly(A+) RNA, Total RNA
• Product Line: Ambion™, MessageAmp™

• Content And 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)
• Reverse Transcriptase: ArrayScript™
• Format: Kit
• Quantity: 20 Reactions