• Detect more genes due to increased Percent Present Calls
  
  
• Obtain sufficient amounts of amplified RNA from a single round of amplification with only 50 ng of input RNA
  
  
• Biotinylated nucleotide included in the optimized NTP mix, reducing set up time and maximizing labeling
  
  
• MEGAscript® IVT reaction ensures optimal representation of the gene expression profile
  
  

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Gene arrays measure the expression levels of thousands of genes in a single experiment. They have become the method of choice for assessing gene expression on a genome-wide scale. The main limitation of this technology, however, is the need for large amounts of RNA for hybridization to the GeneChip®. This is of particular concern when working with limited samples, such as those obtained from laser capture microdissection (LCM), primary cell lines, and mRNA deficient cells and tissues. This limitation is overcome with a T7 linear amplification method commonly referred to as the Eberwine method (Van Gelder et al. 1990). This amplification method produces RNA that most accurately represents the initial mRNA profile and so was the method-of-choice for our MessageAmp™ kits.

The MessageAmp™ II-Biotin Enhanced Kit incorporates extensive improvements to the in vitro transcription (IVT) step of the amplification protocol. The IVT reaction has been optimized for the most effective biotin labeling. An NTP mix containing biotinylated UTP has been added to the kit for ease of use and optimal labeling. These modifications to the kit have significantly increased sensitivity, providing users with increased Percent Present Calls (Figure 2) and sufficient amounts of amplified RNA for array analysis after just a single round of amplification using only 50 ng of input RNA (Figure 3).

The MessageAmp II-Biotin Enhanced Kit includes enough reagents for the single-round amplification and biotin-labeling of 20 samples. A "5X" kit is also available to accomodate 100 reactions. These kits do not include enough reagents for two rounds of amplification. If your experiments require two rounds of amplification, you will need to purchase the MessageAmp II aRNA Amplification Kit (see below) in addition to the MessageAmp II-Biotin Kit.

References
Van Gelder RN, von Zastrow ME, Yool A, Dement WC, Barchas JD, Eberwine JH. (1990) Amplified RNA synthesized from limited quantities of heterogeneous cDNA. Proc Natl Acad Sci USA 87: 1663-7.

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