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规格 |
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| Includes Label or Dye: | Yes |
| Labeling Method: | Direct Labeling |
| Product Line: | Ambion™, TotalPrep™ |
| Reverse Transcriptase: | ArrayScript™ |
| Sample Type: | Poly(A+) RNA, Total RNA |
| System Type: | Illumina Sentrix® |
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储存 |
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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 |
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描述 |
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.
