Illumina® TotalPrep™ RNA Amplification Kit

Catalog number: AMIL1791

Ambion™  Related applications: Microarray Analysis

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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.
For Research Use Only. Not for use in diagnostic procedures.


System: Illumina Sentrix®
Product Size: 24 reactions
Labeling Method: Direct Labeling
Includes Label or Dye: Yes
Reverse Transcriptase: ArrayScript™ Reverse Transcriptase
Sample Type (Specific): Total RNA, Poly(A+) RNA

Contents & storage

The following components should be stored at -20°C in a non-frost-free freezer:
• 26 µl T7 Oligo(dT) Primer
• 51 µl 10X First Strand Buffer
• 205 µl dNTP Mix
• 26 µl RNase Inhibitor
• 27 ArrayScript™ Reverse Transcriptase
• 252 µl 10X Second Strand Buffer
• 51 µl DNA Polymerase
• 26 µl RNase H
• 64 µl T7 10X Reaction Buffer
• 64 µl T7 Enzyme Mix
• 1.75 ml Nuclease-free Water (may be stored at any temperature)
• 10 µl 1 mg/ml Control RNA

Store the following at room temperature:
• 10 ml Nuclease-free Water (may be stored at any temperature)
• 30 ml Wash Buffer (may be stored at 4°C)
• 7 ml cDNA Binding Buffer
• 9 ml cRNA Binding Buffer
• 24 cRNA Filter Cartridges
• 48 cRNA Collection Tubes
• 30 cDNA Filter Cartridges + Tubes
• 30 cDNA Elution Tubes


Manuals & protocols

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