Ambion™ MessageAmp™ Premier RNA Amplification Kit Non-distribution product as customer accommodation. Available on GSA/VA Contract for Federal Government customers only.

Innovative kit for the preparation of RNA samples for microarray analysis

Manufacturer: Invitrogen 4385821

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Catalog No. 43-858-21

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    Product Type Kit, RNA Amplification
    Includes 11μL First Strand Enzyme Mix, 44μL First Strand Buffer Mix, 22μL Second Strand Enzyme Mix, 55μL Second Strand Buffer Mix, 66μL T7 Enzyme Mix, 220μL T7 Biotin IVT Mix, 10μL Control RNA 1mg/mL, 1.75mL Nuclease-free Water, 600μL aRNA Binding Buffer Concentrate, 120μL RNA Binding Beads, 10mL aRNA Wash Solution Concentrate, 5mL aRNA Elution Solution, 1mL 5X Array Fragmentation Buffer, 10 8-Strip PCR Tubes and Caps, 1 U-Bottom Plate, 1 Reservoir, 10mL Nuclease-free Water
    For Use With (Application) Expression Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, RNA Amplification
    Quantity 10 reactions

    Kit employs the proven methodology for RNA sample preparation and labeling based on the original T7 in vitro transcription (IVT) amplification technology, known as the Eberwine or reverse transcription-IVT (RT-IVT) method. The RT-IVT method is considered the gold standard for sample preparation in microarray-based expression profiling. It is well documented in the current scientific literature and was experimentally validated using TaqMan™ RT-PCR. Builds upon the linear RNA amplification method and reagents developed for MessageAmp™ II Kits with enhancements that create a simplified workflow, improved performance, and more flexible RNA input requirements.

    Using the MessageAmp Premier RNA Amplification Kit
    Based on this proven enzymatic amplification and labeling strategy, the MessageAmp Premier procedure starts with a simple cDNA synthesis reaction using ArrayScript™ reverse transcriptase. No cleanup step is required after the RT; cDNA is used directly in a high yield IVT reaction using MEGAscript technology. The IVT is configured to incorporate the modified nucleotide, biotin-UTP, into the aRNA synthesized. (In this protocol, the amplified RNA is referred to as aRNA; in the literature, it is also commonly called cRNA.) Once purified, the aRNA is suitable for use on microarray gene expression analysis platforms designed for biotin-modified antisense RNA samples.

    • Very flexible RNA input requirements: 20-500ng total RNA, depending on the tissue or cell type
    • Streamlined workflow: depending on the RNA input amounts, only a single day of processing time is required to complete a microarray experiment
    • Simplified protocol and streamlined reagents
    • Complete kit: everything needed to prepare biotin-modified aRNA is included
    • Rigorous quality control that includes GeneChip™ analysis
    • T7 MEGAscript™ IVT delivers up to 50,000-fold amplification
    • Single-tube format
    • Easy-to-use, high-recovery, magnetic-bead aRNA purification

    Lowest RNA input requirements for a single round of amplification:

    • 20ng total RNA yields enough biotin-modified aRNA for most microarray formats, with results equivalent to those from samples prepared from significantly more starting material using other RT-IVT kits

    Expression Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, RNA Amplification

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