Description
The Illumina® TotalPrep™ RNA Amplification Kit 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. Experience these improvements:
• Improved first-strand cDNA synthesis using ArrayScript™ reverse transcriptase
• Biotinylated UTP allows ease of use and optimal labeling
• Maximization of cRNA yield using MEGAscript® IVT technology
• Significantly improved sensitivity starting with just 50 ng of input RNA
New improvements for a tested and reliable amplification procedure
The TotalPrep™ RNA Amplification Kit is based on the RNA amplification protocol developed in the laboratory of James Eberwine. The procedure consists of reverse transcription with an oligo(dT) primer bearing a T7 promoter (TTTT-PT7) using ArrayScript™ RT, a proprietary reverse transcriptase (RT) engineered to produce higher yields of first-strand cDNA than wild type enzymes. ArrayScript™ RT catalyzes the synthesis of virtually all 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, the 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