• Perform amplification from just 50 ng of input RNA
  
  
• Developed for expression profiling using the Sentrix® BeadChips
  
  
• Bio-16-UTP now included in optimized NTP mix, reducing set up time and maximizing labeling
  
  

Illumina® TotalPrep™ RNA Amplification Kit
The TotalPrep RNA Amplification Kit (patent pending) is a complete system for generating biotinylated, amplified RNA for hybridization with Illumina Sentrix® arrays. The new TotalPrep RNA Amplification Kit incorporates extensive improvements to the in vitro transcription reaction (IVT) over the previous Illumina RNA Amplification Kit. The IVT reaction has been optimized for the most effective biotin labeling. An NTP mix containing biotinylated UTP has been added to the kit, allowing ease of use and optimal labeling. These modifications to the kit have significantly increased sensitivity, providing increased Percent Present calls and single round amplification with just 50 ng of input RNA.

The TotalPrep RNA Amplification Kit is based on the RNA amplification protocol developed in the laboratory of James Eberwine (Van Gelder et al. 1990). The procedure consists of reverse transcription with an oligo(dT) primer bearing a T7 promoter using Ambion’s proprietary ArrayScript™, a reverse transcriptase (RT) engineered to produce higher yields of first strand cDNA than wild type enzymes. ArrayScript catalyzes the synthesis of virtually full-length cDNA, which is the best way to ensure production of reproducible microarray samples. The cDNA then undergoes second strand synthesis and clean-up to become a template for in vitro transcription with T7 RNA Polymerase. To maximize cRNA yield, Ambion’s proprietary MEGAscript® in vitro transcription (IVT) technology, along with biotin UTP (provided in the kit), is used to generate hundreds to thousands of biotinylated, antisense RNA copies of each mRNA in a sample.

Illumina® TotalPrep™-96 RNA Amplification Kit
The Illumina TotalPrep-96 RNA Amplification Kit is a complete system for generating biotinylated (biotin-16-UTP), amplified RNA for hybridization with Illumina Sentrix® arrays. It is based on the patented Eberwine linear RNA amplification method. The new kit offers magnetic bead-based purification of both the cDNA and the aRNA. The magnetic bead-based purification reduces elution volumes for easier protocol integration. These improvements offer:

• Consistently higher yields with magnetic bead compared to filter based method
• Highly reproducible aRNA yields and size (CV less than 10%)
• Flexible format allows RNA amplification with a partial to a full 96 well plate
• Easily automated for high-throughput amplification of mRNA using robotics